Many types of mammalian cells can aggregate and differentiate into 3-D multicellular spheroids when cultured in suspension or a nonadhesive environment. Compared to conventional monolayer cultures, multicellular spheroids resemble real tissues better in terms of structural and functional properties. Multicellular spheroids formed by transformed cells are widely used as avascular tumor models for metastasis and invasion research and for therapeutic screening. Many primary or progenitor cells on the other hand, show significantly enhanced viability and functional performance when grown as spheroids. Multicellular spheroids in this aspect are ideal building units for tissue reconstruction. Here we review the current understanding of multicellular spheroid formation mechanisms, their biomedical applications, and recent advances in spheroid culture, manipulation, and analysis techniques. ...

Multicellular tumor spheroid cultures (MCTS), have a wide variety of uses in the field of cancer treatment. Current methods, however, do not provide spheroids adequate large for therapy testing. In order to circumvent this problem, a bioreactor made using Polydimethylsiloxane (PDMS) was constructed to allow the adequate growth of spheroid clusters. Liver Hepatic Carcinoma cells (Hep G2) and Anaplastic Thyroid Carcinoma (SW 1736) were cultured and isolated. They were then further matured using the hanging drop method, involving spheroid formation using gravity. The optimal growth time for hanging drop cultures was determined to be 72 hours. PDMS wells of different diameter were then constructed using a 24-well plate as a base, and clusters of cells were transferred into them for MCTS formation. The wells were fabricated using PDMS as a mold, then carving out wells for cell growth. Development of the spheroids in the bioreactor was monitored using microscopy paired with various staining ...

The Cultrex® 3D Spheroid Fluorometric Proliferation/Viability Assay utilizes a 3D Culture Qualified 96 Well Spheroid Formation Plate alongside a specialized Spheroid Formation ECM to drive aggregation and/or spheroid formation of cells. Upon completion of spheroid formation, the spheroid may be treated with pharmacological agents to evaluate tumor viability after drug treatment. Tumor spheroid expansion is visualized microscopically and can be quantitated through image analysis software for real-time and label free evaluation. At the conclusion of the assay, cell viability may be assessed by fluorescence using Resazurin. The 3D Spheroid Fluorometric Proliferation/Viability Assay offers an in vitro, standardized, three-dimensional, high content format for inducing multicellular tumor spheroid (MCTS) formation and quantifying cell viability within the spheroids in response to pharmacological treatment.. ...

We have evaluated the sensitivity to immunotoxins (IT) of monolayer and of 200-250 microns multicellular tumor spheroid (MTS) cultures obtained with human breast (MCF7) and glioblastoma (U118) tumor cells and with rat glioblastoma (9L) cells. Monolayer MCF7 and U118 cells were highly sensitive to antitransferrin receptor (anti-TfnR) ricin A chain (RTA)-IT (Tfn-RTA and MAb OKT9-RTA) treatment in the presence of the intracellular RTA-IT enhancing agent human serum albumin-monensin (HSA-Mo) conjugate. A 790- to 2000-fold higher concentration of anti-TfnR IT was instead required to reduce by 50% the volume of individually treated MCF7 spheroids, as evaluated by applying the Gompertz growth model. Monolayer 9L cells showed 230- to 5700-fold lower sensitivity to Tfn-RTA IT than MCF7 and U118 monolayers, yet 9L spheroid cells were almost as sensitive to anti-TfnR IT as monolayer 9L cultures. Binding studies performed with [125I]-Tfn and FITC-labelled anti-TfnR MAb revealed that 9L monolayers and MTS ...

VitroGel 3D-Advanced hydrogel for 3D cell culture and spheroid culture research. We offer many matrices and other solutions for 3D cell and spheroid culture

TY - JOUR. T1 - Cisplatin resistant spheroids model clinically relevant survival mechanisms in ovarian tumors. AU - Chowanadisai, Winyoo. AU - Messerli, Shanta M.. AU - Miller, Daniel H.. AU - Medina, Jamie E.. AU - Hamilton, Joshua W.. AU - Messerli, Mark A.. AU - Brodsky, Alexander S.. N1 - Funding Information: This work was funded by the National Institutes of Health (NIH) NCRR supplement grant P41 RR001395-27S1 (JWH), National Science Foundation (NSF) DBI-1005378 ?REU Site: Biological Discovery in Woods Hole?, faculty startup funds from the Office of Research at Oklahoma State University (WC), and the Mary Kay Foundation (ASB). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Joel Commisso at the Interdisciplinary Center for Plasma Mass Spectrometry at the University of California, Davis, for his technical expertise. We thank Milton C. Gum for his advice regarding dataset analyses. This work was funded by ...

The Cellartis DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium paired with a perfusion bioreactor generates large amounts of high-quality, hiPS cells.

he livers role in metabolism of chemicals makes it an appropriate tissue for toxicity testing. Current testing protocols, such as animal testing and two-dimensional liver cell systems, offer limited resemblance to in vivo liver cell behaviour, in terms of gene expression profiles and metabolic competence; thus, they do not always accurately predict human toxicology. In vitro three-dimensional liver cell models offer an attractive alternative. This study reports on the development of a 3D liver model, using HepG2 cells, by a hanging-drop technique, with a focus on evaluating spheroid growth characteristics and suitability for genotoxicity testing. The cytokinesis-blocked micronucleus assay protocol was adapted to enable micronucleus (MN) detection in the 3D spheroid models. This involved evaluating the difference between hanging vs non-hanging drop positions for dosing of the test agents and comparison of automated Metafer scoring with manual scoring for MN detection in HepG2 spheroids. The ...

Hierarchically Assembled Mesenchymal Stem Cell Spheroids Using Biomimicking Nanofilaments and Microstructured Scaffolds for Vascularized Adipose Tissue

Spheroids vs. Organoids-Whats the difference?. Spheroids and organoids are both 3D structures made of many cells. Although the terminology has been used interchangeably, there are distinct differences between the two. An organoid is a collection of organ-specific cell types that develops from stem cells or organ progenitors, which self-organize through cell sorting and spatially restricted lineage commitment in a manner similar to in vivo.1 Multicellular tumor spheroid models were first described in the early 1970s and were obtained by culturing cancer cell lines under non-adherent conditions which forces cells to adhere to each other to form spheroids. A substantial difference between these 3D structures is the higher order of self-assembly and dependence on an extracellular matrix for organoids as opposed to spheroid cultures.1 Tissue-derived tumor spheres and organotypic multicellular spheroids are typically obtained by tumor tissue mechanical dissociation and tumorosphere cultures can be ...

RGD (arginine-glycine-aspartic) containing elastin-like proteins (RGD-ELP), in the structure of TGPG[VGRGD (VGVPG)6]20WPC, was treated to HEK293 cells to construct in vivo-mimic 3-dimensional (3D) spheroid cells. The constructed 3D cells had increased extracellular matrices (ECMs). To identify the induced functions of 3D cells with ECMs, genomic profiles changed in 3D cells compared to the conventional 2-dimensional (2D) cells were analyzed. In HEK293 cells exposed RGD-ELP for 72 h, the total up- and down-regulated genes (824 genes) were selected to be differentially expressed genes. Further analysis by bioinformatics showed that top functions induced in 3D cells were cell proliferation, cell-to-cell signaling, cell differentiation and movement, and connective tissue development and function. We propose that the 3D cells constructed with RGD-ELP in this study could provide more in vivo-mimic responses by cell-to-cell and cell-to-ECM signaling. © 2013 Korean Society of Environmental Risk ...

Multicellular tumor spheroids are models of increasing interest for cancer and cell biology studies. They allow considering cellular interactions in exploring cell cycle and cell division mechanisms. However, 3D imaging of cell division in living spheroids is technically challenging and has never been reported. Here, we report a major breakthrough based on the engineering of multicellular tumor spheroids expressing an histone H2B fluorescent nuclear reporter protein, and specifically designed sample holders to monitor live cell division dynamics in 3D large spheroids using an home-made selective-plane illumination microscope. As illustrated using the antimitotic drug, paclitaxel, this technological advance paves the way for studies of the dynamics of cell divion processes in 3D and more generally for the investigation of tumor cell population biology in integrated system as the spheroid model.

Three-dimensional (3D) cancer cell cultures are being utilized for drug screening with increasing frequency since they provide a more physiologically relevant solid tumor model than two-dimensional cell cultures. Spheroids are a well characterized, in vitro tumor model system that has been utilized with multiple cancer cell lines. Growth and manipulation of these cultures in hanging drop plates (HDPs) can be challenging to manipulate and these challenges are amplified as sample throughput increases. We automated the culture and compound treatment of cancer spheroids in HDPs, as well as the preparation of these spheroids for analysis by high content imaging and flow cytometry. Automated cancer spheroid cultures showed excellent consistency in size and shape, providing higher confidence in the screening results. Creating single cell suspensions from the cancer spheroids for flow cytometry analysis was another significant challenge overcome through automation. Imaging provided information such as ...

Epithelial ovarian cancer (EOC) has a relatively high mortality rate (~55%). One of the presiding causes is that the current chemotherapeutic regimes are unable to achieve sustained remission, despite frequently producing a positive response at first treatment. One of the reasons that EOC is difficult to treat is that the mechanism of dissemination is unusual. EOC dissemination characteristically involves local invasion of pelvic and abdominal organs. Unlike many epithelial cancers, initial dissemination rarely requires the vasculature, although the vasculature is often implicated in the advanced stages of disease. Recently, it has become apparent that aggregates of malignant cells (spheroids) contained within malignant ascites represent a significant impediment to efficacious treatment of late stage EOC. In vivo, spheroids are present in the malignant ascites of EOC patients, while in vitro cultured spheroids are capable of tumorgenesis in vivo and display a reduced response to chemotherapeutic ...

TY - JOUR. T1 - Application of Concave Microwells to Pancreatic Tumor Spheroids Enabling Anticancer Drug Evaluation in a Clinically Relevant Drug Resistance Model. AU - Yeon, Sang Eun. AU - No, Da Yoon. AU - Lee, Sang Hoon. AU - Nam, Suk Woo. AU - Oh, Il Hoan. AU - Lee, Jaehwi. AU - Kuh, Hyo Jeong. PY - 2013/9/10. Y1 - 2013/9/10. N2 - Intrinsic drug resistance of pancreatic ductal adenocarcinoma (PDAC) warrants studies using models that are more clinically relevant for identifying novel resistance mechanisms as well as for drug development. Tumor spheroids (TS) mimic in vivo tumor conditions associated with multicellular resistance and represent a promising model for efficient drug screening, however, pancreatic cancer cells often fail to form spheroids using conventional methods such as liquid overlay. This study describes the induction of TS of human pancreatic cancer cells (Panc-1, Aspc-1, Capan-2) in concave polydimethylsiloxane (PDMS) microwell plates and evaluation of their usefulness as ...

Abstract. Tumor microenvironments present significant barriers to penetration by antibodies and immunoconjugates and are difficult to study in vitro. Cells cultured as monolayers typically exhibit less resistance to therapy than those grown in vivo. Therefore, it is important to develop an alternative research model that better represents in vivo tumors. We have developed a protocol to produce multicellular spheroids, a simple and more relevant model of in vivo tumors that allows for further investigations of the microenvironmental effects on drug penetration and tumor cell killing. The protocol is used to produce in vitro three-dimensional tumor spheroids from established human cancer cell lines and primary cancer cells isolated from patients without the use of any extracellular components. To study the ability of tumor-targeting immunoconjugates to penetrate these tumor spheroids in vitro, we have used an immunotoxin targeting mesothelin, a surface protein expressed in malignant mesotheliomas. ...

Learn how AggreWell™ Microwell Plates work to generate large numbers of uniform, size-controlled 3D cell culture spheroids for many areas of research, including cancer, stem cells, drug discovery and more

With their novel and proprietary design, these microplates are ideal for generating and analyzing 3D multicellular spheroids in the same microplate. The Ultra-Low Attachment surface enables uniform and reproducible 3D multicellular spheroid formation. The black opaque microplate body shields each optically clear, round bottom well from well-to-well crosstalk.

BACKGROUND AIMS: Mesenchymal stromal cells (MSCs) exhibit the inherent potential to regulate multiple signaling pathways and cell types that contribute to the pathogenesis of inflammatory and immune diseases. However, more recent studies have suggested that the secretion of immunomodulatory factors by MSCs can be enhanced by three-dimensional aggregation or pro-inflammatory cytokine treatment.. METHODS: Human MSC spheroids were formed by forced aggregation into agarose micro-wells and subsequently cultured in either minimal essential medium alpha supplemented with fetal bovine serum or serum-free, defined MesenCult-XF medium (STEMCELL Technologies, Vancouver, Canada). A subset of the spheroids were treated with pro-inflammatory cytokines interferon (IFN)-γ or tumor necrosis factor (TNF)-α or both for 4 days. Immunomodulatory factor (prostaglandin E2, indoleamine 2,3-dioxygenase, transforming growth factor-β1 and interleukin-6) secretion was quantified after 4 days of culture, and the ...

Multicellular tumor spheroids are used as a model to assess the efficacy of replicating oncolytic adenoviruses. As most assays used to assess cellular viability are unsuitable for oncolytic viruses because of ongoing viral replication, we have used positron emission tomography (PET) to sequentially determine the incorporation of 18F-labeled deoxyglucose ( 18F-DG) as a measure of viability and compared the results to more commonly used assays for measuring the effect of oncolytic therapy. Glioma monolayer cultures and spheroids were infected with wild-type replicating adenovirus and viability was measured by 18F-DG incorporation, WST-1 assay, crystal violet assay, and spheroid volume 2 to 10 days following infection. Results show that volume measurements in adenovirus-infected spheroids are confounded by the cytopathic effect occurring in infected cells. 18F-DG PET provides a useful method to assess small differences in cell number and viability following oncolytic viral therapy in glioma ...

View Cultrex 3-D Spheroid Basement Membrane Extract Cell Invasion Assay, 96-well, Cell Culture Products for Basement Membrane Extracts from R&D Systems.

Peritoneal dissemination is a critical prognostic factor in ovarian cancer. Although stabilized spheroid formation promotes cancer cell peritoneal dissemination in ovarian cancer, the associated oncogenes are unknown. In this study, we assessed the role of the KRAS oncogene in ovarian cancer cell dissemination, focusing on the stability of cells in spheroid condition, as well as the modulation of intracellular signaling following spheroid transformation. We used ID8, a murine ovarian cancer cell line, and ID8-KRAS, an oncogenic KRAS (G12 V)-transduced ID8 cell line in this study. Spheroid-forming (3D) culture and cell proliferation assays were performed to evaluate the growth characteristics of these cells. cDNA microarray analysis was performed to identify genes involved in KRAS-associated signal transduction in floating condition. A MEK inhibitor was used to evaluate the effect on cancer peritoneal dissemination. Cell viability and proliferation in monolayer (2D) cultures did not differ between ID8

Peritoneal dissemination is a critical prognostic factor in ovarian cancer. Although stabilized spheroid formation promotes cancer cell peritoneal dissemination in ovarian cancer, the associated oncogenes are unknown. In this study, we assessed the role of the KRAS oncogene in ovarian cancer cell dissemination, focusing on the stability of cells in spheroid condition, as well as the modulation of intracellular signaling following spheroid transformation. We used ID8, a murine ovarian cancer cell line, and ID8-KRAS, an oncogenic KRAS (G12 V)-transduced ID8 cell line in this study. Spheroid-forming (3D) culture and cell proliferation assays were performed to evaluate the growth characteristics of these cells. cDNA microarray analysis was performed to identify genes involved in KRAS-associated signal transduction in floating condition. A MEK inhibitor was used to evaluate the effect on cancer peritoneal dissemination. Cell viability and proliferation in monolayer (2D) cultures did not differ between ID8

The use of transition metal based tumour-activated prodrugs is a promising strategy to improve the selectivity of anticancer agents towards cancer over healthy tissues. The rational design of these agents is difficult, as knowledge and tools to understand their biological activities have been lacking to date. In this study, multiple fluorescent cellular models have been developed and characterised to study metal complexes [M(acac)3] and [M(dbm)3], as models of hypoxia-activated prodrugs, where M = Co, Fe, Ru. The biological activities of these metal complexes were evaluated in both the monolayer cultures as well as spheroids, an exemplary in vitro tumour model. As most anticancer agents have been developed with the purpose of damaging the nucleus and the DNA, the changes in the cell cycle progression caused by the prodrugs was investigated using the fluorescent ubiquitination cell cycle indicator (FUCCI) system. When the metal complexes chaperone the cytotoxin to the hypoxic region of the ...

The pharmaceutical actions of DNG on the expression of aromatase and COX-2 and the production of PGE2 were examined using spheroid cultures of human ESCs. More aromatase, COX-2, and PGE2 were expressed in spheroid cultures than in conventional ESCs monolayers. In the spheroid cultures, DNG (10−7 M) and progesterone (10−7 M) inhibited the expression of aromatase, COX-2, and PGE2. DNG also inhibited NF-κB DNA-binding activity and reduced the immunocytochemical protein expression of aromatase, COX-2, and NF-κB p50 nuclear localization.. Conclusion(s): ...

HER2 is overexpressed in 20% invasive breast tumors and correlates with low free disease survival. Trastuzumab (Tz), monoclonal antibody anti HER2, is used to treat HER2+ tumors; however more than half of the patients are resistant or acquire resistance during treatment. Multicellular tumor spheroids are a 3D cell growth model that mimics the structure of in vivo avascular tumors, with heterogenic cell subpopulations developed due to differential oxygen and nutrient supply through the spheroid. We have previously demonstrated that HER2+ cells cultured as spheroids are more resistant to Tz than monolayers. We also observed that in spheroids Tz inhibited basal apoptosis and was capable of inducing autophagy, leading to Tz resistance. In addition, cancer stem cells (CSC), widely associated with chemotherapy resistance, were specifically targeted by Tz.. The aim of this study was to analyze the resistance acquired in 3D and the impact of the CSC developed in these conditions.. Since Tz-treated BT474 ...

Doxorubicin-mediated radiosensitivity in multicellular spheroids from a lung cancer cell line is enhanced by composite micelle encapsulation Wen-Hong Xu,1 Min Han,2 Qi Dong,3 Zhi-Xuan Fu,3 Yuan-Yuan Diao,2 Hai Liu,3 Jing Xu,3 Hong-Liang Jiang,4 Su-Zhan Zhang,3 Shu Zheng,3 Jian-Qing Gao,2 Qi-Chun Wei11Department of Radiation Oncology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, 2Institute of Pharmaceutics, College of Pharmaceutical Sciences, 3Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, School of Medicine, 4Department of Polymer Science and Engineering, Zhejiang University, Hangzhou, Zhejiang, ChinaBackground: The purpose of this study is to evaluate the efficacy of composite doxorubicin-loaded micelles for enhancing doxorubicin radiosensitivity in multicellular spheroids from a non-small cell lung cancer cell line.Methods: A novel composite doxorubicin-loaded micelle consisting

Chronic alcohol intake leads to neuroinflammation and astrocyte dysfunction, proposed to perpetuate alcohol consumption and to promote conditioned relapse-like binge drinking. In the present study, human mesenchymal stem cells (MSCs) were cultured in 3D-conditions to generate MSC-spheroids, which greatly increased MSCs anti-inflammatory ability and reduced cell volume by 90% versus conventionally 2D-cultured MSCs, enabling their intravenous administration and access to the brain. It is shown, in an animal model of chronic ethanol intake and relapse-drinking, that both the intravenous and intra-cerebroventricular administration of a single dose of MSC-spheroids inhibited chronic ethanol intake and relapse-like drinking by 80-90%, displaying significant effects over 3-5 weeks. The MSC-spheroid administration fully normalized alcohol-induced neuroinflammation, as shown by a reduced astrocyte activation, and markedly increased the levels of the astrocyte Na-glutamate (GLT-1) transporter. This research

Author(s): Davey, Shruti Krishna | Advisor(s): Varghese, Shyni | Abstract: Techniques for cellular encapsulation within three-dimensional (3D) structures, such as bioprinting and patterning methods, play an important role in creating complex and hierarchically organized tissues, as well as when studying cell-cell and cell-matrix interactions. To this end, advances in technologies have enabled development of methods to generate such 3D structures. We describe an easy-to-use photopatterning method involving photomask and a simple fluorescence microscope. This method is adapted to generate homogeneous and co-culture tissue constructs. Additionally, we extend this approach to establish a system to quantitatively study cancer spheroid growth. We developed a method combining the photomask-based 3D photopatterning technique with microfluidics technology to encapsulate a cancer spheroid within a patterned hydrogel embedded with fluorescent particles, monitor the cancer growth, and quantify the corresponding

Developing a new drug from bench to bedside requires up to 10 years and investments between 0,8 and 1,1 billion US Dollars, according to the recent published data by the US Food and Drug Administration (FDA). As many as 90% of new drug candidates fail during the clinical development stage. New assays predicting the efficacy of drugs in the early, pre-clinical stage and which prevent animal testing are required to reduce both the development time and the costs in drug development. Modern assays are performed with three-dimensional cell cultures. They essentially avoid hard and flat surfaces. Hence, they favor less constrained physiological sample dynamics and promote cells growth under tissue-like conditions. This improves the reliability and the physiological significance of the assay. We develop three-dimensional cultures of tumor cell spheroids for phenotypic drug screening issues. The cultivated spheroids resemble their microenvironment and serve as a model for tumors in the living organism. ...

Tumor spheroids are becoming an important tool for the investigation of malignancy stem cell (CSC) function in tumors; therefore low-cost and high-throughput methods for drug testing of tumor spheroids are needed. screening of a panel of anti-proliferative medicines to assess inhibitory effects on the Citalopram Hydrobromide growth Mouse monoclonal to CD16.COC16 reacts with human CD16, a 50-65 kDa Fcg receptor IIIa (FcgRIII), expressed on NK cells, monocytes/macrophages and granulocytes. It is a human NK cell associated antigen. CD16 is a low affinity receptor for IgG which functions in phagocytosis and ADCC, as well as in signal transduction and NK cell activation. The CD16 blocks the binding of soluble immune complexes to granulocytes. of malignancy stem cells in 3-D ethnicities. Keywords: neurospheres tumor spheroids cancers stem cell glioblastoma acridine orange microscopy Solid tumors develop within a three-dimensional (3-D) spatial conformation which isnt mimicked by two-dimensional (2-D) ...

Tumor spheroids are becoming an important tool for the investigation of malignancy stem cell (CSC) function in tumors; therefore low-cost and high-throughput methods for drug testing of tumor spheroids are needed. screening of a panel of anti-proliferative medicines to assess inhibitory effects on the Citalopram Hydrobromide growth Mouse monoclonal to CD16.COC16 reacts with human CD16, a 50-65 kDa Fcg receptor IIIa (FcgRIII), expressed on NK cells, monocytes/macrophages and granulocytes. It is a human NK cell associated antigen. CD16 is a low affinity receptor for IgG which functions in phagocytosis and ADCC, as well as in signal transduction and NK cell activation. The CD16 blocks the binding of soluble immune complexes to granulocytes. of malignancy stem cells in 3-D ethnicities. Keywords: neurospheres tumor spheroids cancers stem cell glioblastoma acridine orange microscopy Solid tumors develop within a three-dimensional (3-D) spatial conformation which isnt mimicked by two-dimensional (2-D) ...

Prostate cancer metastasis: roles of recruitment and reprogramming, cell signal network and three-dimensional growth characteristics

Nanoscale hydroxyapatite (HA) is an optimal candidate biomaterial for bone tissue engineering because of its bioactive and osteoconductive properties. In this study, micro- and nanoscale HA particles with rod- and wirelike morphology were synthesized by a novel sol-gel-hydrothermal process. Sol-gel chemistry was used to produce a dry gel containing amorphous calcium phosphate (ACP), which was used as a precursor material in a hydrothermal process. The sol-gel-hydrothermal products were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) to determine particle morphology, crystal structure, and the presence of chemical functional groups. A pure HA crystal was synthesized, which underwent both one- and three-dimensional growth, resulting in tunable microrod and nanorod, and wire morphologies. The effects of solution pH and reaction time on particle diameter and length were assessed. Particle diameter ranged from 25 to 800 nm and

TY - JOUR. T1 - Niche-localized tumor cells are protected from HER2-targeted therapy via upregulation of an anti-apoptotic program in vivo. AU - Zoeller, Jason J.. AU - Bronson, Roderick T.. AU - Selfors, Laura M.. AU - Mills, Gordon B.. AU - Brugge, Joan S.. PY - 2017/12/1. Y1 - 2017/12/1. N2 - Several lines of evidence suggest that components of the tumor microenvironment, specifically basement membrane and extracellular matrix proteins, influence drug sensitivities. We previously reported differential drug sensitivity of tumor cells localized adjacent to laminin-rich extracellular matrix in three-dimensional tumor spheroid cultures. To evaluate whether differential intratumor responses to targeted therapy occur in vivo, we examined the sensitivity of human epidermal growth factor receptor 2-positive tumors to lapatinib using a previously described ductal carcinoma in situ-like model characterized by tumor cell confinement within ductal structures surrounded by an organized basement membrane. ...

Drug development in pancreatic cancer has lacked advances seen in other solid organ cancers. Self-renewing CSCs undergoing asymmetric division within their niche are able to drive pancreatic cancer progression and mediate resistance to chemo-, molecular, and radiotherapy (7, 21, 34). The presence of these progenitor-like cells correlates with poor clinical outcomes of survival and tumor recurrence (23, 35, 36). In an attempt to address these challenges, we examined how spheroid culture models responded to drug treatments relative to traditional monolayer cultures. Submitting the PANC1 cell line to HTS-enabled drug screening in both spheroid and monolayer formats provided a relative assessment of drugs with preferential or equal cell-killing phenotype in both formats (20, 27). A limited number of approved or investigational drugs demonstrated similar capacities to kill pancreatic cancer spheroids relative to monolayer cultures. Among those was a previously reported inhibitor of ITK (NCGC00188382; ...

|em|The Scientist |/em|is bringing together a panel of experts to discuss the value of spheroid culture systems, and to explore the technical benefits and challenges of making the switch from 2-D to 3-D culture.

My research interests are focused on liver metabolism and function as well as hepatic pathologies. We previously established an integrated 3D spheroid cell culture system for primary human hepatocytes (PHH) in which cells remain viable and functionally stable for multiple weeks. Importantly, we recently showed that PHH in this model exhibited superior sensitivity to hepatotoxic agents compared to other emerging cell models, such as HepG2 and HepaRG cells, and were faithfully reproducing in vivo drug toxicity mechanisms in man. Furthermore, using a combination of untargeted and targeted metabolomics, we showed that the endogenous and xenobiotic metabolic signatures of PHH were maintained in 3D spheroids, thus allowing to comprehensively study drug-induced molecular effects on cellular metabolism and to investigate mechanisms of drug action. The results indicate that the 3D PHH spheroid system faithfully mimics heptic phenotypes in vivo and can be utilized for long-term analyses of drug ...

PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

Personalized Medicine Based Approach to Model Patterns of Chemoresistance and Tumor Recurrence Using Ovarian Cancer Stem Cell Spheroids In order to screen the effect of chemotherapeutics and biologics on resistant ovarian cancers with a personalized basis, investigators developed a 3D hanging drop spheroid platform. They initiated spheroids with primary ALDH+ CD133+ ovarian CSCs from different patient samples, and demonstrated that stem cell progeny from harvested spheroids was similar to the primary tumor. [Clin Cancer Res] Abstract S100A4 Is a Biomarker and Regulator of Glioma Stem Cells that Is Critical for Mesenchymal Transition in Glioblastoma Scientists showed that S100A4 is a novel biomarker of glioma stem cells (GSCs). Selective ablation of S100A4-expressing cells was sufficient to block tumor growth in vitro and in vivo. They also identified S100A4 as a critical regulator of GSC self-renewal in mouse and patient-derived glioma tumorspheres. [Cancer Res] Abstract Metformin Targets ...

Microtissues offers a 12 well spheroids pack containing: three 12-256 micro-molds for making small spheroids and three 12-81 micro-molds for making larger spheroids. Embryoid bodies mimic early in vivo development. Multicellular tumor spheroids mimic the in vivo tumor environment.. ...

Fallopian tube secretory epithelial cells (FTSECs) have been implicated as a cell-of-origin for high-grade serous epithelial ovarian cancer. However, there are relatively few in vitro models of this tissue type available for use in studies of FTSEC biology and malignant transformation. In vitro three-dimensional (3D) cell culture models aim to recreate the architecture and geometry of tissues in vivo and restore the complex network of cell-cell/cell-matrix interactions that occur throughout the surface of the cell membrane. We have established and characterized 3D spheroid culture models of primary FTSECs. FTSEC spheroids contain central cores of hyaline matrix surrounded by mono- or multi-layer epithelial sheets. We found that 3D culturing alters the molecular characteristics of FTSECs compared to 2D cultures of the same cells. Gene expression profiling identified more than a thousand differentially expressed genes between 3D and 2D cultures of the same FTSEC lines. Pathways significantly under

A primary aim of cancer therapy is to irradicate solid tumor masses. Anticancer drugs are nevertheless not systematically developed with this aim in mind ‐ screening and mechanistic studies are usually performed using cells grown under 2‐D monolayer conditions. We have developed a procedure to screen for compounds that induce apoptosis of 3‐D multicellular tumor spheroids. The chemical properties of spheroid screening hits differed significantly from conventional monolayer screening hits with regard to compound hydrophobicity and polar surface area. The spheroid screen resulted in hits which were more effective than standard chemotherapeutical agents in inducing wide‐spread apoptosis of tumor spheroids. Hypotheses regarding the mechanisms of action for the selected compounds were generated using the Connectivity Map (CMAP; a compendium of gene expression signatures from drug treated cell lines). One of the hits in the spheroid screen was the alkaloid thaspine from the South American tree ...

Porvair Sciences reports, due to promising results, it has agreed with its collaborative partners to make additional investment in the CEAT project** which aims to dramatically improve the diagnosis and treatment of ovarian cancer.. In the initial stage of the CEAT (Cluster for Epigenomic and Antibody Drug Conjugate Therapeutics) project - Porvair Sciences supplied its proprietary Chromatrap® bead-free Chromatin Immunoprecipitation (ChIP) technology to help University of Swansea researchers develop new epigenomic profiling approaches to enable advances in drug development and patient profiling.. So far the project has had success in developing bespoke chemo-resistant OC cell lines, optimised the Chromatrap® kits for 3D spheroid analysis along with biochemical analysis on AFM (Atomoic Force Microscopy). The expansion to the project will allow current microrheology assessment of disease phenotype spheroid culture models and high resolution binding kinetics to support selective targeting and drug ...

Nutrient regulation using continuous growth adjusted feeding improves growth rates of mammalian cell spheroids compared to intermittent ...

The advent of 3D printing technologies promises to make microfluidic organ-on-chip technologies more accessible for the biological research community. To date, hydrogel-encapsulated cells have been successfully incorporated into 3D printed microfluidic devices. However, there is currently no 3D printed microfluidic device that can support multicellular spheroid culture, which facilitates extensive cell-cell contacts important for recapitulating many multicellular functional biological structures. Here, we report a first instance of fabricating a 3D printed microfluidic cell culture device capable of directly immobilizing and maintaining the viability and functionality of 3D multicellular spheroids. We evaluated the feasibility of two common 3D printing technologies i.e. stereolithography (SLA) and PolyJet printing, and found that SLA could prototype a device comprising of cell immobilizing micro-structures that were housed within a microfluidic network with higher fidelity. We have also ...

article{8536566, abstract = {Multicellular tumor spheroid models serve as an important three-dimensional in vitro cell model system as they mimic the complex tumor micro-environment and thus have contributed to valuable assays in drug discovery studies. In this study, we present a state-of-the-art laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) setup for high spatial resolution elemental imaging of multicellular tumor spheroids and an approach to account for variations in cell density. A low dispersion LA-ICP-MS setup was employed, providing accelerated throughput, high sensitivity and permitting a lateral image resolution down to {\texttildelow}2.5 {\textmu}m for phosphorus and platinum in HCT116 colon cancer spheroids upon treatment with the clinically used anti-cancer drug oxaliplatin. Phosphorus was introduced as scalar to compensate for differences in cell density and tissue thickness and the Pt/P ratios together with the high resolution adopted in our approach ...

TY - JOUR. T1 - Multicellular spheroids from normal and neoplastic thyroid tissues as a suitable model to test the effects of multikinase inhibitors. AU - Cirello, Valentina. AU - Vaira, Valentina. AU - Grassi, Elisa Stellaria. AU - Vezzoli, Valeria. AU - Ricca, Dario. AU - Colombo, Carla. AU - Bosari, Silvano. AU - Vicentini, Leonardo. AU - Persani, Luca. AU - Ferrero, Stefano. AU - Fugazzola, Laura. PY - 2017/2/7. Y1 - 2017/2/7. N2 - Multicellular three-dimensional (3D) spheroids represent an experimental model that is intermediate in its complexity between monolayer cultures and patients tumor. In the present study, we characterize multicellular spheroids from papillary (PTC) and follicular (FTC) thyroid cancers and from the corresponding normal tissues. We show that these 3D structures well recapitulate the features of the original tissues, in either the differentiated and stem-like components. As a second step, we were aimed to test the effects of a small multikinase inhibitor, SP600125 ...

Hereditary diffuse leukoencephalopathy with spheroids (HDLS) is a rare adult onset autosomal dominant disorder characterized by cerebral white matter degeneration with demyelination and axonal spheroids leading to progressive cognitive and motor dysfunction. Spheroids are axonal swellings with discontinuous or absence of myelin sheaths. It is believed that the disease arises from primary microglial dysfunction that leads to secondary disruption of axonal integrity, neuroaxonal damage, and focal axonal spheroids leading to demyelination. Spheroids in HDLS resemble to some extent those produced by shear stress in a closed head injury with damage to axons, causing them to swell due to blockage of axoplasmic transport. In addition to trauma, axonal spheroids can be found in aged brain, stroke, and in other degenerative diseases. In HDLS, it is uncertain whether demyelination occurs prior to the axonal spheroids or what triggers neurodegeneration after apparently normal brain and white matter ...

Video articles in JoVE include Studying Surfactant Effects on Hydrate Crystallization at Oil-Water Interfaces using a Low-Cost Integrated Modular Peltier Device, In Vitro Model of Human Cutaneous Hypertrophic Scar using Macromolecular Crowding, Potato Virus X-based microRNA Silencing (VbMS) In Potato., Performing Colonoscopic-Guided Pinch Biopsies in Mice and Evaluating Subsequent Tissue Changes, Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification, Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection, Application of Atomic Force Microscopy to Detect Early Osteoarthritis, A 3D Spheroid Model for Glioblastoma, A Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) Platform for Investigating Peptide Biosynthetic Enzymes, Using a Chemical Biopsy for Graft Quality Assessment.

Tumor hypoxia is associated clinically with therapeutic resistance and poor patient outcomes. One feature of tumor hypoxia is activated expression of carbonic anhydrase IX (CA9), a regulator of pH and tumor growth. In this study, we investigated the hypothesis that impeding the reuptake of bicarbonate produced extracellularly by CA9 could exacerbate the intracellular acidity produced by hypoxic conditions, perhaps compromising cell growth and viability as a result. In 8 of 10 cancer cell lines, we found that hypoxia induced the expression of at least one bicarbonate transporter. The most robust and frequent inductions were of the sodium-driven bicarbonate transporters SLC4A4 and SLC4A9, which rely upon both HIF1α and HIF2α activity for their expression. In cancer cell spheroids, SLC4A4 or SLC4A9 disruption by either genetic or pharmaceutical approaches acidified intracellular pH and reduced cell growth. Furthermore, treatment of spheroids with S0859, a small-molecule inhibitor of sodium-driven

Tumor hypoxia is associated clinically with therapeutic resistance and poor patient outcomes. One feature of tumor hypoxia is activated expression of carbonic anhydrase IX (CA9), a regulator of pH and tumor growth. In this study, we investigated the hypothesis that impeding the reuptake of bicarbonate produced extracellularly by CA9 could exacerbate the intracellular acidity produced by hypoxic conditions, perhaps compromising cell growth and viability as a result. In 8 of 10 cancer cell lines, we found that hypoxia induced the expression of at least one bicarbonate transporter. The most robust and frequent inductions were of the sodium-driven bicarbonate transporters SLC4A4 and SLC4A9, which rely upon both HIF1α and HIF2α activity for their expression. In cancer cell spheroids, SLC4A4 or SLC4A9 disruption by either genetic or pharmaceutical approaches acidified intracellular pH and reduced cell growth. Furthermore, treatment of spheroids with S0859, a small-molecule inhibitor of sodium-driven

TY - JOUR. T1 - Cardiosphere-derived cells from pediatric end-stage heart failure patients have enhanced functional activity due to the heat shock response regulating the secretome. AU - Kaushal, Sunjay. AU - Sharma, Sudhish. AU - Mishra, Rachana. AU - Simpson, David L.. AU - Wehman, Brody. AU - Colletti, Evan J.. AU - Deshmukh, Savitha. AU - Datla, Srinivasa Raju. AU - Balachandran, Keerti. AU - Guo, Yin. AU - Chen, Ling. AU - Siddiqui, Osama T.. AU - Kaushal, Shalesh. PY - 2015/4/1. Y1 - 2015/4/1. N2 - We have demonstrated that human neonatal cardiosphere-derived cells (CDCs) derived from the young are more regenerative due to their robust secretome. However, it is unclear how the decompensated pediatric heart impacts the functional activity of their CDCs. Our aim was to characterize the potency of pediatric CDCs derived from normal functioning myocardium of control heart disease (CHD) patients to those generated from age-matched end stage heart failure (ESHF) patients and to determine the ...

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Evodiamine (EVO) and rutaecarpine (RUT) are promising anti-tumor drug candidates. The evaluation of the anti-proliferative activity and cellular uptake of EVO and RUT in 3D multicellular spheroids of cancer cells would better recapitulate the native situation and thus better reflect an in vivo response to the treatment. Herein, we employed the 3D culture of MCF-7 and SMMC-7721 cells based on hanging drop method and evaluated the anti-proliferative activity and cellular uptake of EVO and RUT in 3D multicellular spheroids, and compared the results with those obtained from 2D monolayers. The drugs IC50 values were significantly increased from the range of 6.4-44.1 μM in 2D monolayers to 21.8-138.0 μM in 3D multicellular spheroids, which may be due to enhanced mass barrier and reduced drug penetration in 3D models. The fluorescence of EVO and RUT was measured via fluorescence spectroscopy and the cellular uptake of both drugs was characterized in 2D tumor models. The results showed that the cellular

Curcumin is a polyphenolic compound derived from Curcumin longa L. There is growing body of data showing the antitumor effect of curcumin in different cancers; however, the molecular mechanism underlying of this inhibition in breast cancer is still remained to be elucidated. Here we investigated the antitumor activity of curcumin alone or in combination with paclitaxel or doxorubicin in MCF-7 cells in monolayer cell cultures and spheroids models. Moreover, the cytotoxic activity of three different forms of curcumin (phytosomal), phospholipidated curcumin, amorphous curcumin and turmeric oleoresin were evaluated, compared to unformulated curcumin. ...

A Polymer Thin Film Platform that Promotes Direct Conversion of Cancer Cell Lines to Tumorigenic Cell Spheroids. Yu, Seung Jung; Choi, Minsuk; Choi, Yoonjung; Lee, Eun-beol; Lee, Eunjung; Kim, Jinyong; Kang, Sukmo; et al, 2018 BMES Annual Meeting, Biomedical Engineering Society, 2018-10- ...

The aim of the present study was to assess the feasibility of a 3D tumor cell culture model, that is, multicellular tumor spheroids (MCTSs) as an adequate model for micrometastases and therefore as a pharmacological model for efficacy testing of trifunctional therapeutic antibodies. Unlike conventio …

Anoikis resistance is a key to the survival of cells in malignant transformation and metastasis [18, 19]. It may also be a key to the adaptation of cells to suspension culture and spheroids growth used in engineering. For epithelial cells, suppression of anoikis upon detachment seems to be induced when cell-cell contacts are formed. For example, cadherin-mediated homotypic interactions maintain the survival of human prostate carcinoma DU-145 cells in the absence of extracellular matrix (ECM) attachments [20]. Also, disruption of E-cadherin cell-cell contacts showed more important for suppressing anoikis of normal enterocytes after detachment from villus epithelium, as compared to cell-ECM disruption [17]. Growth as spheroids renders tumor cells less sensitive to exogenous apoptotic stimuli, and spheroids have greater drug resistance than the corresponding monolayers [21]; and some cells used in engineering also grow as spheroids in suspension. Thus, elucidating the mechanisms by which spheroids ...

Tindall, M. J. and Dyson, L and Smallbone, K. and Maini, P. K. (2012) Modelling acidosis and the cell cycle in multicellular tumour spheroids. Journal of Theoretical Biology, 298 (1). pp. 107-115. Smallbone, K. and Maini, P. K. and Gatenby, R. A. (2010) Episodic, transient systemic acidosis delays evolution of the malignant phenotype: Possible mechanism for cancer prevention by increased physical activity. Biology Direct, 5 (22). pp. 1-8. Maini, P. K. and Gatenby, R. A. and Smallbone, K. (2010) Modelling of tumour metabolism. In: Proceedings of the International Congress of Mathematicians, Hyderbad, 19th-27th August. Hindustan Book Agency, New Delhi, pp. 3091-3104. Smallbone, K. and Gatenby, R. A. and Maini, P. K. (2008) Mathematical modelling of tumour acidity. Journal of Theoretical Biology, 255 (1). pp. 106-112. Gatenby, R. A. and Smallbone, K. and Maini, P. K. and Rose, F. and Averill, J. and Nagle, R. B. and Worrall, L. and Gillies, R. J. (2007) Cellular adaptations to hypoxia and acidosis ...

Real-time monitoring of cisplatin cytotoxicity on three-dimensional spheroid tumor cells NamHuk Baek,1,* Ok Won Seo,1,* Jaehwa Lee,1 John Hulme,2 Seong Soo A An2 1Department of Research and Development, NanoEntek Inc., Seoul, 2Department of BioNano Technology, Gachon University, Gyeonggi-do, Korea *These authors contributed equally to this work Abstract: Three-dimensional (3D) cell cultivation is a powerful technique for monitoring and understanding diverse cellular mechanisms in developmental cancer and neuronal biology, tissue engineering, and drug development. 3D systems could relate better to in vivo models than two-dimensional (2D) cultures. Several factors, such as cell type, survival rate, proliferation rate, and gene and protein expression patterns, determine whether a particular cell line can be adapted to a 3D system. The 3D system may overcome some of the limitations of 2D cultures in terms of cell-cell communication and cell networks, which are essential for understanding differentiation,

To circumvent this problem here we adopted SP cells followed by the spheroid culture technique, to enrich cancer stem-like cells. We performed a chemical

To investigate the effects of age and disease on endogenous cardiac progenitor cells, we obtained right atrial and left ventricular epicardial biopsies from patients (n = 22) with chronic ischaemic heart disease and measured doubling time and surface marker expression in explant- and cardiosphere-derived cells (EDCs, CDCs). EDCs could be expanded from all atrial biopsy samples, but sufficient cells for cardiosphere culture were obtained from only 8 of 22 ventricular biopsies. EDCs from both atrium and ventricle contained a higher proportion of c-kit+ cells than CDCs, which contained few such cells. There was wide variation in expression of CD90 (atrial CDCs 5-92 % CD90+; ventricular CDCs 11-89 % CD90+), with atrial CDCs cultured from diabetic patients (n = 4) containing 1.6-fold more CD90+ cells than those from non-diabetic patients (n = 18). No effect of age or other co-morbidities was detected. Thus, CDCs from atrial biopsies may vary in their therapeutic potential.

Tumor colonization involves reciprocal interactions between the metastatic cancer cells and various stromal cells in foreign microenvironment. Fibroblasts are the second most numerous cell types in omentum, which is the commonest site of ovarian cancer metastasis. However, the crosstalk between metastatic ovarian cancer cells and fibroblasts that involved in tumor colonization remains largely unknown. In this study, we used a three-dimensional (3D) co-culture model to study the role of normal fibroblasts on tumor colonization of ovarian cancer cells. Human ovarian cancer cell lines, which were established from peritoneal ascites, were seeded on normal fibroblasts embedded in extracellular matrix (Matrigel). Our results showed that ovarian cancer spheroids formed in Matrigel with fibroblasts were significantly larger and also more invasive than those formed in Matrigel without fibroblasts, suggesting the cancer-fibroblast interaction promotes the colonization of ovarian cancer cells. To search ...

Precise cell count and viability determination of adipose-derived stem cells, hematopoietic stem cells, microcarrier cultures, cells in spheroid cultures

The forces that multicellular tumor aggregates exert on their environment lead to non-linear, scale-invariant tissue deformations far away from the tumor, which can be exploited to quantify its collective contractility.

Background: To improve current treatment strategies for patients with aggressive colorectal cancer (CRC), the molecular understanding of subgroups of CRC with poor prognosis is of vast importance. SOX2 positive tumors have been associated with a poor patient outcome, but the functional role of SOX2 in CRC patient prognosis is still unclear. Methods: An in vitro cell culture model expressing SOX2 was used to investigate the functional role of SOX2 in CRC. In vitro findings were verified using RNA from fresh frozen tumor tissue or immunohistochemistry on formalin fixed paraffin embedded (FFPE) tumor tissue from a cohort of 445 CRC patients. Results: Using our in vitro model, we found that SOX2 expressing cells displayed several characteristics of cancer stem cells; such as a decreased proliferative rate, a spheroid growth pattern, and increased expression of stem cell markers CD24 and CD44. Cells expressing SOX2 also showed down-regulated expression of the intestinal epithelial marker CDX2. We ...

50 µCi quantities of [G-3H]-Vinblastine Sulphate are available for your research. Application of [3H]Vinblastine can be found in: human placental transport and the contribution of P-glycoprotein (P-gp) in pharmacology, oocytes expressing human P-gp (probing trans- and cis-inhibitory effects on efflux) in pharmacology, accumulation of the antimalarial microtubule inhibitors by a major human malarial parasite in biochemical pharmacology, diffusivity and distribution in three-dimensional tumour tissue (experimental and mathematical modelling) in cancer research, etc. ...

Purpose : Mesenchymal stem cells (MSCs) have been shown to be a potential treatment for degenerative eye diseases. In this study we aim to identify and compare levels of signaling factors secreted by MSCs cultured in 2D monolayer (2D-MSCs) or in spheroids (spheoird MSCs). To explore potential non cell-based therapies, we also treated retinal ganglion cells (RGC-5) with MSC derived microvesicles (MVs) and analyzed levels of secreted signaling factors.. Methods : We seeded 2D-MSCs, spheroid MSCs and cells isolated by trypsinization of spheoird MSCs at equal cell density and collected conditioned medium after 24hrs. Hanging drop protocol was used to prepare spheroid MSCs. We isolated microvesicles (MVs) from conditioned medium of MSCs cultured in those three conditions and co-cultured them with RGC-5 for different time period. Levels of 51 cytokines in conditioned medium of MSCs or MV treated RGC-5 were qualified with a high throughput bead-based assay.. Results : We found that IL-8, IL-6 and CXCL1 ...

The 96 Well 3D Spheroid BME Cell Invasion Assay offers a standardized, three dimensional, high content format for quantitating the degree to which invasive cells penetrate a barrier, consisting of basement membrane components, in vitro in response to chemoattractants and/or inhibiting compounds, which is fundamental for angiogenesis1, embryonic development2, immune response3, and tumor cell metastasis4. Basement membranes are continuous sheets of specialized extracellular matrix that form an interface between endothelial, epithelial, muscle, or neuronal cells and their adjacent stroma. They not only support cells and cell layers, but they also play an essential role in tissue organization that influences cell adhesion, migration, proliferation, and differentiation. Basement membranes are degraded and regenerated during development and wound repair, and they are major barriers to invasion by metastatic tumor cells. Current methods for assessing cell invasion through basement membrane barrier most ...

Primary human hepatocytes (PHH) are commonly used for in vitro studies of drug-induced liver injury. However, when cultured as 2D monolayers, PHH lose crucial hepatic functions within hours. This dedifferentiation can be ameliorated when PHH are cultured in sandwich configuration (2Dsw), particularly when cultures are regularly re-overlaid with extracellular matrix, or as 3D spheroids. In this study, the six participating laboratories evaluated the robustness of these two model systems made from cryopreserved PHH from the same donors considering both inter-donor and inter-lab variability and compared their suitability for use in repeated-dose toxicity studies using 5 different hepatotoxins with different toxicity mechanisms ...

All keratocytes participate in sphere formation. A: Freshly isolated primary keratocytes were cultured in polyHEMA-coated dishes either in Advanced-DMEM (ADV, d

Cell polarity is essential for cells to divide asymmetrically, form spatially restricted subcellular structures and participate in three-dimensional multicellular organization. PAR proteins are conserved polarity regulators that function by generating cortical landmarks that establish dynamic asymmetries in the distribution of effector proteins. Here, we review recent findings on the role of PAR proteins in cell polarity in C. elegans and Drosophila, and emphasize the links that exist between PAR networks and cytoskeletal proteins that both regulate PAR protein localization and act as downstream effectors to elaborate polarity within the cell.. ...

Here, a protocol to quantify epithelial early common progenitor/stem cells grown as spheres in non‐adherent culture conditions is described

3D Cell Culture is much better at replicating in vivo environment than traditional two-dimensional cultures. Learn about the history of spheroids.

Overview Automation compatible - Secure handling - No surface attachment The world famous and unique hanging drop plate in 96-well format with automation-compatible top-loading. Loading of cell suspension from the top through unique SureDrop™ inlet engineered for highly uniform and stable hanging drops Gravity-assist

Wound healing is a complex process that requires an interplay between several cell types. Classically, fibroblasts have been viewed as producers of extracellular matrix, but more recently they have been recognized as orchestrators of the healing response, promoting and directing, inflammation and neovascularization processes. Compared to those from healthy tissue, inflammation-associated fibroblasts display a dramatically altered phenotype and have been described as sentinel cells, able to switch to an immunoregulatory profile on cue. However, the activation mechanism still remains largely uncharacterized. Nemosis is a model for stromal fibroblast activation. When normal human primary fibroblasts are deprived of growth support they cluster, forming multicellular spheroids. Clustering results in upregulation of proinflammatory markers such as cyclooxygenase-2 and secretion of prostaglandins, proteinases, cytokines, and growth factors. Fibroblasts in nemosis induce wound healing and tumorigenic ...

Claudia Fischbach of Cornell University and associates create stunning pictures, such because the one right here, utilizing a brand new collagen-embedded multicellular spheroid platform. The research explores the connections between tissue microenvironment and weight problems, which can have vital implications for breast most cancers malignancy in overweight sufferers.. At its most extreme, this way of thinking results in eco-fascism, the belief the hurt people do to Earth can be reduced by chopping the number of non-white individuals. What the were the virus meme tells us about green politics.As Covid-19 pressured people into lockdown, memes emerged showing the earth was therapeutic due to our absence. These were false claims - however their recognition revealed how seductive the damaging idea that were the virus may be. The shifting wall represents the time interval between the last concern available in JSTOR and essentially the most recently revealed issue of a journal.. In uncommon ...

Full-Length Fibronectin Drives Fibroblast Accumulation at the Surface of Collagen Microtissues during Cell-Induced Tissue Morphogenesis. on Wyss Institute

Supplementary MaterialsAdditional file 1: Physique S1. inhibitors targeting CSCs from the ethyl acetate (EtOAc) extract of the roots of and to evaluate their in vitro anti-cancer activities. Methods The chemical components of the EtOAc extract and the subfractions of were isolated by using various column chromatographies on silical gel, Sephadex LH-20, and preparative HPLC. Their chemical structures were then decided on the basis of spectroscopic data including NMR, MS and IR analysis and their physicochemical properties. The inhibitory effects of the isolated compounds against STAT3 signaling were screened by a STAT3-dependent luciferase reporter gene assay. The tyrosine phosphorylation of STAT3 was examined by Western Blot analysis. In vitro anti-cancer effects of the STAT3 pathway inhibitor were further evaluated on cell growth of human being HCC cells by a MTT assay, on self-renewal capacity of HCC CSCs from the tumorsphere formation assay, and on cell cycle and apoptosis by circulation ...

Experimental data, particularly derived from tumour spheroids, indicate that drug penetration barriers may be an important determinant of cytotoxic drug efficacy, even in spheroids of only a few hundred microns in diameter. Clinically, tumour masses of this size would equate with those micrometastas …

In this contribution, an algorithm is presented for counting cells in the first-, second- and third-layers of three-dimensional three-layer cell clusters f

Background: The existence of cancer stem cells (CSCs) or cancer stem-like cells in a tumor mass is believed to be responsible for tumor recurrence because of their intrinsic and extrinsic drug-resistance characteristics ...