Herpes simplex keratitis (HSK) is a sight threatening ocular infection often requiring a specific and prompt laboratory diagnosis. Isolation of Herpes simplex virus (HSV-1) in culture provides the most reliable and specific method and is considered as the Gold Standard in the laboratory diagnosis of HSK in spite of its low sensitivity. Using cell lines of corneal origin for virus isolation may be beneficial under such circumstances, since these cells have been shown to be excellent substrates for the growth of HSV-1 isolated from the cornea. We report a comparative study of a novel human corneal epithelial cell line (HCE) and the Vero cell line in the isolation of HSV-1 from corneal scrapings employing a shell vial assay. Corneal scrapings were obtained from 17 patients with a clinical diagnosis of HSK. All the cases were confirmed by virological investigations (PCR and viral antigen detection positive, n = 15, PCR positive, n = 1, Viral antigen positive, n = 1). Scrapings obtained from 10 patients
TY - JOUR. T1 - Growing human corneal epithelium on collagen shield and subsequent transfer to denuded cornea in vitro. AU - He, Yu Guang. AU - McCulley, James P.. PY - 1991/1/1. Y1 - 1991/1/1. N2 - Three fundamental in vitro experiments have been done in the present report: 1) comparison of three different nutrient media on their abilities to culture and passage the human corneal epithelial cells; 2) evaluation of the ability of extracellular matrix material to promote the growth of cultured human corneal epithelium on collagen corneal shields; and 3) determination of the feasibility of the shield to serve as a carrier for the transfer of cultured cells to allogeneic, denuded corneal surface in vitro. Primary cultures of human corneal epithelium were established from explants which were obtained from limbal and peripheral corneal tissue by three different nutrient media respectively: KGM (Keratinocyte Growth Medium), SHEM (Supplemental Hormonal Epithelial Medium), and one combination of the two ...
Abstract: : Purpose: Proteinase-activated receptors (PARs) are a recently described, novel family of G-protein-coupled receptors, which are activated by the cleavage of their N-terminal domain by serine proteases. These receptors are activated by serine proteases such as trypsin and thrombin released after tissue inflammation and injury. In this study we determined if human corneal cells express functional PAR-1 and PAR-2 receptors and examined the effects of receptor activation on corneal proinflammatory cytokine production. Methods: Human primary corneal epithelial cells (HCEC) and the human corneal epithelial cell line HCE-T were cultured to 80% confluence and then exposed to 10 nM thrombin, 100 M human PAR-1 peptide agonist TFLLRN-NH2, 10 nM trypsin, and 100 M murine PAR-2 peptide agonist SLIGRL-NH2 for 3, 6 and 24 hours. Cells treated with PMA (50ng/ml) and TNF-α-treated (10ng/ml) cells served as positive controls for these studies. PAR-1 and PAR-2 mRNA levels were determined by RT-PCR and ...
Purpose: Autologous serum (AS) eye drops offer a potential treatment alternative for non-healing corneal epithelial defects in clinical practice. In corneal epithelial cell cultures, fetal bovine serum (FBS) is often used to support the growth of the cells. The dose-dependent effect of AS and FBS on viability, migration and proliferation of human corneal epithelial cells (HCECs) have not been specified yet. The purpose of this study was to analyse the concentration-dependent effects of AS and FBS on HCEC viability, migration and proliferation, in vitro.. Methods: First, AS was prepared from 13 patients according to the regulations of the LIONS Cornea Bank Saar-Lor-Lux, Trier/Westpfalz. HCECs were firstly cultured in DMEM/F12 with 5% FBS, 0.5% DMSO, 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media which was consisting of DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS for 24 hours. Thereafter, HCEC viability was analysed using Cell ...
Rabbit Corneal Epithelial Cells from Creative Bioarray are isolated from corneal tissue of New Zealand White Rabbit. Rabbit Corneal Epithelial Cells are grown in a T25 tissue culture flask pre-coated with gelatin-based coating solution for 2 min and incubated in Creative Bioarrays Culture Complete Growth Medium for 3-5 days. Cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 0.5x10^6 cells per ml and is delivered frozen. Cells can be expanded for 3-7 passages at a split ratio of 1:2 under the cell culture conditions specified by Creative Bioarray. Repeated freezing and thawing of cells is not recommended ...
TY - JOUR. T1 - Induced expression of insulin-like growth factor-1 by amniotic membrane-conditioned medium in cultured human corneal epithelial cells. AU - Lee, Joon H.. AU - Ik, Hee Ryu. AU - Kim, Eungkweon. AU - Lee, Jongeun. AU - Hong, Soon Won. AU - Hyung, Keun Lee. PY - 2006/3/1. Y1 - 2006/3/1. N2 - PURPOSE. To determine the effect of amniotic membrane-conditioned medium (AMCM), via insulin-like growth factor (IGF)-1 induction, on human corneal epithelial cell (HCEC) proliferation. METHODS. HCECs were cultured from corneal limbal tissue with supplemented hormonal epithelial medium (SHEM). After administration of AMCM, cell proliferation was evaluated with an MTT assay and DNA synthesis with methyl-[3H]-thymidine incorporation assay. RT-PCR and Western immunoblot analyses were performed, to determine potential inducible factors that may be associated with AMCM-induced cell proliferation. Neutralizing anti-IGF-1 antibody and small interfering (si)RNA were also used to clarify the role of ...
Full Text - Corneal transparency, dependent on the integrity of epithelial cells, is essential for vision. Corneal epithelial damage is one of the most commonly observed ocular conditions and proper wound healing is necessary for corneal transparency. Sirt6, a histone deacetylase, has been shown to regulate many cellular events including aging and inflammation. However, its specific role in corneal epithelial wound healing remains unknown. Here we demonstrated that Sirt6 was expressed in corneal epithelial cells and its expression decreased with age. In an in vivo corneal epithelial wound healing model, Sirt6 deficiency resulted in delayed and incomplete wound healing and was associated excessive inflammation in the corneal stroma and dysfunction of Notch signaling, leading to keratinization of the corneal epithelium and corneal opacity. Aging Sirt6-deficient mice spontaneously developed corneal keratitis with extensive infiltration of inflammatory cells into the cornea. In vitro
Corneal Epithelial Cell Growth Kit (ATCC ® PCS-700-040) contains components that when added to Corneal Epithelial Cell Basal Medium (ATCC ® PCS-700-030) create a complete ATCC ® Primary Cell Solutions ™ culture environment for corneal epithelial cells derived from normal human cornea (e.g., Primary Corneal Epithelial Cells, Normal, Human, ATCC ® PCS-700-010). The serum-free medium formulation is designed to support normal corneal cell morphology as well as promote rapid growth and proliferation. No feeder layers, extracellular matrix proteins or other substrates are required.
Purpose: Our previous studies show that human corneal epithelial cells (HCEC) have a functional vitamin D receptor (VDR) and respond to vitamin D by dampening TLR-induced inflammation. Here, we further examined the timing of the cytokine response to combined vitamin D-TLR treatment and used genome-wide microarray analysis to examine the effect of vitamin D on corneal gene expression.. Methods: Telomerase-immortalized HCEC (hTCEpi) were stimulated with polyinosinic-polycytidylic acid (poly[I:C]) and 1,25-dihydroxyvitamin D3 (1,25D3) for 2 to 24 hours and interleukin (IL)-8 expression was examined by quantitative (q)PCR and ELISA. Telomerase-immortalized HCEC and SV40-HCEC were treated with 1,25D3 and used in genome-wide microarray analysis. Expression of target genes was validated using qPCR in both cell lines and primary HCEC. For confirmation of IκBα protein, hTCEpi were treated with 1,25D3 for 24 hours and cell lysates used in an ELISA.. Results: Treatment with 1,25D3 increased ...
The healthy corneal epithelium has tight junctions and forms a barrier between the environment and the inside of the eye. The tear film aids in protecting the corneal surface, and artificial tear...
TY - JOUR. T1 - Osmotic tolerance of rabbit corneal epithelium in tissue culture. AU - Dale, R. Meyer. AU - McCulley, James P. PY - 1991. Y1 - 1991. N2 - The osmotic tolerance of stratified rabbit corneal epithelial cultures was determined for a balanced salt solution and various pharmaceutical vehicles in an effort to determine the biocompatibility of the different formulations after 4-128 min exposures. Acute damage was assessed by monitoring the release of radiolabeled nucleotides and corresponding changes in tissue morphology of the prelabeled cultures. Protracted damage was evaluated by re-examining the morphology of the same cultures after 24 hr along with quantitative measurements of residual 3H release, tissue loss, and concomitant changes in protein synthesis. Our testing results indicated that the permissive limits of a balanced salt solution were between 305 and 400 mOsm, which is comparable to the guidelines for native epithelium in situ established by organ perfusion studies. In ...
In order to validate in situ corneal redox fluorometry, the redox state and phosphorylation potential of freeze trapped rabbit corneal epithelium and endothelium were studied using quantitative histochemical methods. The results were compared with noninvasive measurements using an optically sectioning fluorometer microscope. Enucleated rabbit eyes were either frozen in Freon-12, cooled by liquid nitrogen or exposed for 1 hr in 1 mM NaCN to block oxidation and then freeze trapped. Corneas were sectioned, freeze-dried, samples of individual layers dissected, weighed, and analyzed for: NADH, NAD+, NADPH, NADP+, ATP, ADP, and Pi. The aerobic epithelium showed a ratio for NAD+/NADH of 1.85 +/- 0.08 (9). In anoxia this ratio decreased to 1.06 +/- 0.07 (8). The NAD+/NADH ratio of aerobic endothelium was 3.25 +/- 0.28 (6); in anoxia this ratio was 0.68 +/- 0.14 (5). The values of phosphorylation potential ATP/(ADP X Pi)M-1 were: 447.9 +/- 40.2 (9) in aerobic epithelium, 378.2 +/- 24.7 (5) in anoxic epithelium;
Invest Ophthalmol Vis Sci. 2014 Mar 20;55(3):1770-9. doi: 10.1167/iovs.13-12988. The Epstein-Barr virus causes epithelial-mesenchymal transition in human corneal epithelial cells via Syk/src and Akt/Erk signaling pathways. Park GB1, Kim D, Kim YS, Kim S, Lee HK, Yang JW, Hur DY. Author information Department of Anatomy and Research Center for Tumor Immunolo...
Tan S-S, Williams EA, Tam PPL: X-chromosome inactivation occurs at different times in different tissues of the post-implantation mouse embryo. Nat Genet 1993, 3:170-174. Roach SA. (1968). The theory of random clumping. London: Methuen. Collinson, J. M., Morris, L., Reid, A. I., Ramaesh, T., Keighren, M. A., Flockhart, J. H., Hill, R. E., Tan, S.-S., Ramaesh, K., Dhillon, B. and West, J. D. (2002). Clonal analysis of patterns of growth, stem cell activity and cell movement during the development and maintenance of the murine corneal epithelium. Dev. Dyn. 224: 432-440. Mort, R. L., Ramaesh, T. Kleinjan, D, A., Morley, S. D., and West. J. D. (2009). Mosaic analysis of stem cell function and wound healing in the mouse corneal epithelium. BMC Dev Bio 9:4. West, J. D. (1975). A theoretical approach to the relation between patch size and clone size in chimaeric tissue. J. Theor. Biol. 50, 153-160. West JD. (1976). Clonal development of the retinal epithelium in mouse chimaeras and X-inactivation ...
Ultraviolet (UV)-induced cataracts are becoming a major environmental health concern because of the possible decrease in the stratospheric ozone layer. Experiments were designed to isolate gene(s) affected by UV irradiation in rabbit cornea tissues using fluorescent differential display-reverse transcription-polymerase chain reaction (FDDRT-PCR). The epithelial cells were grown in standard medium for 2 or 4 hours post treatment. Cornea epithelial cells were irradiated with UVB for 20 minutes. RNA was extracted and amplified by reverse transcriptase-polymerase chain reaction using poly A+ specific anchoring primers and random arbitrary primers. Polyacrylamide gel electrophoresis revealed several differentially expressed genes in untreated versus UV irradiated cells. Complimentary DNA (cDNA) fragments resulting from fluorescent differentially expressed mRNAs were eluted from the gel and re-amplified. The re-amplified PCR products were cloned directly into the PCR-TRAP cloning system. These data showed
Purpose. Damage to the corneal epithelium results in the massive secretion of fibronectin (FN) shortly after injury and induces the expression of its integrin receptor α5β1. The authors reported previously that FN induces α5 expression in human corneal epithelial cells and rabbit corneal epithelial cells by altering the binding of the transcription factor (TF) Sp1 to a regulatory element from the α5 promoter that it is also flanked by binding sites for the TFs NFI and AP-1. Here, they assessed the function of NFI and AP-1 on α5 gene expression and evaluated the contribution of FN to their overall regulatory influence. Methods. TF binding to the α5 promoter was evaluated in vitro by electrophoretic mobility shift assays and in vivo by ligation-mediated PCR or chromatin immunoprecipitation. TFs expression was monitored by Western blot, whereas their influence was assessed by transfection and RNAi analyses. Results. Coexpression of Sp1, NFI, and AP-1 was demonstrated in all cell types, and ...
PURPOSE: To evaluate the effects of fluoroquinolone-based antibacterial ophthalmic solutions on cell proliferation in vitro and corneal wound healing in vivo. METHODS: Statens Serum institute rabbit corneal cells were exposed to phosphate-buffered saline, 1.5% and 0.5% levofloxacin, 0.5% moxifloxacin, and 0.3% gatifloxacin, for 2 min, following which the cells were incubated without the drug. The cell viability was evaluated after 24 or 72 h of incubation. Rabbit corneal epithelial abrasion models created using n-heptanol were instilled with saline or fluoroquinolone-based solutions 7 times at 30-min intervals, following which corneal epithelial wound healing was evaluated from 30 min to 48 h by the measurement of electrical corneal resistance (CR) ratios ...
This sounds like an ulcer with an attitude, but it really refers to the way the epithelium rolls up on itself at the edge of the crater, instead of covering across the top of it. The treatment is to numb the eye, debride the edge with a sterile swab, and perform a keratotomy. With a tiny needle, the veterinarian makes a series of cuts/scratches in a grid pattern over the ulcer; lines crossing from the normal corneal epithelium to the ulcerated across to the normal epithelium on the other side. It seems counterintuitive to "scratch" the injured eye, but it leaves a path for the cells to grow across ...
Figure 2. Localization of PMCA in human corneal epithelium. A: Immunostaining with pan-PMCA antibody (5F10) revealed strong PMCA labeling in all layers of the corneal epithelium (CE). The staining was associated primarily with the plasma membranes. The stroma (S) was unstained. B: Control section incubated with nonimmune mouse IgG was unstained.. ...
A primary culture of normal corneal epithelium was immortalized with Ad12-SV40 hybrid virus by overnight incubation with virus at 37°C
antibody-antibodies.com is the marketplace for research antibodies. Find the right antibody for your research needs. Rapamycin Prolongs the Survival of Corneal Epithelial Cells in Culture.
If you have a question about this talk, please contact .. Abstract not available. This talk is part of the Foster Talks series.. ...
OBJECTIVE To explore the possibility of using telomerase as a marker of corneal limbal stem cells. METHODS Corneal limbal tissues and central corneal epithelial tissues from 8 rabbits were examined for telomerase activity qualitatively by telomere repeat amplification protocol (TRAP) and quantitatively by detecting the light value with bioluminescent technique. 5-Fu (20 mg) was injected subconjunctivally in 4 rabbits and the other 4 rabbits were not injected. RESULTS Telomerase activity was positive in all corneal limbal tissues and negative in all central corneal tissues. Telomerase activity of corneal limbal cells (light value 165,575) was significantly higher than that of central corneal epithelial cells (light value 34,912) by bioluminescent technique (P = 0.001). 5-Fu injection group showed higher telomerase activity than that of the group without injection (light value 145,754) (P = 0.02). CONCLUSIONS Positive telomerase activity is detected in the corneal limbal tissues. It suggests that
PURPOSE. A previous report has described an ocular surface reconstruction method involving the use of cultivated corneal epithelium derived from limbal explants. In the current study, a new culture system was developed involving the in vitro propagation on amniotic membrane (AM) of epithelial cells from enzymatically dissociated limbal epithelium. The purpose of this new method is to produce a cultivated epithelial cell layer that contains stem cells and that is superior to explanted cultivated epithelium. The new cell-suspension technique was compared with the existing explant method. METHODS. Limbal epithelial cells were dissociated from donor eyes by dispase and seeded on the denuded AM. Small pieces of limbal epithelium were also cultured on denuded AM as explant cultures. The cultivated epithelium was examined by electron microscopy and immunohistochemistry for cornea-specific keratins (K3 and K12). RESULTS. Both cell-suspension and explant culture methods produced a healthy epithelial cell ...
Cultured limbal tissue transplants have become widely used over the last decade as a treatment for limbal stem cell deficiency (LSCD). While the number of patients afflicted with LSCD in Australia and New Zealand is considered to be relatively low, the impact of this disease on quality of life is so severe that the potential efficacy of cultured transplants has necessitated investigation. We presently review the basic biology and experimental strategies associated with the use of cultured limbal tissue transplants in Australia and New Zealand. In doing so, we aim to encourage informed discussion on the issues required to advance the use of cultured limbal transplants in Australia and New Zealand. Moreover, we propose that a collaborative network could be established to maintain access to the technology in conjunction with a number of other existing and emerging treatments for eye diseases. ...
Limbal tissue was harvested from cadavers within 12 hours of biological death. The conjunctiva was incised and separated from the limbal junction. A small rectangular shape limbal graft was dissected away and towards the cornea. The graft was cultured on lens capsule cells in corneal epithelial cell culture medium, comprised of Dulbecco-modified Eagles medium (DMEM) supplemented with human AB serum (20%, human serum Type AB), L-glutamine ( 200 mM/mL), penicillin (10,000 U/mL), and streptomycin (10 mg/mL).. ...
Topical insulin application has been proved recently to increase corneal reepithelization rate over diabetic animals. However, its effectiveness on corneal epithelial wound healing in patients who received pars planar vitrectomy (PPV) for diabetic retinopathy and penetrating keratoplasty has not been reported. In this study, we plan to perform a prospective randomized study to determine the effectiveness of topical insulin as a primary treatment for corneal epithelial defect in patients undergoing vitrectomy for diabetic retinopathy and penetrating keratoplasty. All patients enrolled in this study have received corneal epithelial debridement at the end of the ocular surgeries, namely PPV for diabetic retinopathy and penetrating keratoplasty. The patients were randomized into two treatment groups. In the control group, the patients receive conventional postoperative eye drops including topical steroid, antibiotic and mydriatics. In the experimental group, the patients receive topical insulin eye ...
Fungal keratitis is a kind of intractable and sight-threatening diseases. Spleen-tyrosine kinase (Syk) is a non-receptor tyrosine kinase, which plays an important role in the signaling pathway of the receptors. In the current study, we investigate the expression and function of Syk in human corneal epithelial cells with Aspergillus fumigatus (A. fumigatus) infection. Cultured telomerase-immortalized human corneal epithelial cells (THCEs) were treated with A. fumigatus hyphae with or without treatment of Syk inhibitors. Activation of Syk and the role of Syk in regulating inflammatory cytokines and chemokines expression were evaluated. The mRNA expression was determined by real time PCR, and protein activation was measured by western blotting. Syk protein was detected in THCEs, and its activation was enhanced after treatment of A. fumigatus hyphae. Expression of inflammatory cytokines (IL-1β and IL-6) and chemokines (IL-8 and CXCL1) mRNA were significantly increased after stimulation of A. fumigatus
Corneal epithelial thickness has been found to remodel and change significantly after femto-LASIK, according to the findings of a prospective study conducted under the direction of A. John Kanellopoulos, MD, and colleagues. ...
In the present study, we used AM-based limbal explant culture to explore the possible roles of HGF and KGF on cell proliferation and differentiation. AM is a natural matrix substrate, which has been widely used for aiding the repair of corneal epithelium in experimental animals and in patients with corneal epithelial defects (Tseng et al., 1998; Meller and Tseng, 2000; Tseng et al., 2002; Ti et al., 2004). AM-based limbal explant culture has proved to be a good culture system to expand limbal epithelial cells to form limbal-like epithelia equivalents (Wang et al., 2003). The AM-based limbal epithelial equivalents have been successfully used to reconstruct the surface of limbal-deficient cornea, implying that the primitive corneal epithelial cells or possibly corneal stem cells are probably preserved and expanded upon AM-based limbal explant culture (Tsai et al., 2000). Because of its successful clinical application, it is therefore of cell biological and clinical significance to study the ...
Cell therapy is an emerging therapeutic strategy aimed at replacing or repairing severely damaged tissues with cultured cells. Specifically, ocular burns cause depletion of limbal stem cells, which leads to corneal opacification and visual loss. Corneal stem cells are segregated in the basal layer of the limbus, which is the transitional zone of the epithelium located between the cornea and the bulbar conjunctiva. Autologous cultured limbal epithelial cells can restore damaged corneas. We sought to establish a culture system that allows preservation of limbal stem cells and preparation of manageable epithelial sheets. We outline some quality criteria, which assure the clinical performance of keratinocyte culture: evaluation of the number of holoclones within a cultured epithelial graft, proportion of aborting colonies, and percentage of cells expressing high levels of ΔNp63α ...
A 66-years-old male was referred for decreased vision in the left eye. He had a history of previous episodes of keratitis treated several times without improvement. Best- corrected visual acuity was 20/20 in the right eye and 60/200 in the left eye. Slit-lamp examination of the left eye revealed a geographic, gray, translucent, and well- demarcated lesion strictly limited to the epithelium. No associated stromal abnormalities were seen. Ocular examination results of the right eye were unremarkable. Corneal scrapping with histopathologic examination of the removed tissue disclosed intraepithelial neoplasia. The patient underwent corneal epithelial debridement. Post-operative treatment with topical mitomycine C and Ciclosporine A was prescribed, resulting in a complete resolution of the corneal lesion ...
The outermost layer of the cornea consists of a stratified non-keratinized pseudosquamous epithelium known as corneal epithelium (CE). Its highly ordered architecture must be precisely maintained during homeostasis and re-established upon mechanical lesions in order to ensure clear vision. During corneal maturation, the basal epithelium stratifies resulting in differentiated suprabasal layers (wing cells) and more superficial cells that are ultimately eliminated through exfoliation (Thoft and Friend, 1983). At homeostasis the CE undergoes intensive cell renewal. Limbal stem cells localized at the periphery of the cornea undergo slow cell cycle and generate a population of fast-dividing cells called transient amplifying cells (TACs) (Cotsarelis et al., 1989). TACs migrate towards the center of the cornea and populate the basal epithelium to replenish the loss of suprabasal cells that are exfoliating from the corneal surface (Cotsarelis et al., 1989; Davanger and Evensen, 1971). Despite this ...
Journal of Immunology Research is a peer-reviewed, Open Access journal that provides a platform for scientists and clinicians working in different areas of immunology and therapy. The journal publishes research articles, review articles, as well as clinical studies related to classical immunology, molecular immunology, clinical immunology, cancer immunology, transplantation immunology, immune pathology, immunodeficiency, autoimmune diseases, immune disorders, and immunotherapy.
TY - JOUR. T1 - The tumour-suppressor scribble dictates cell polarity during directed epithelial migration: regulation of Rho GTPase recruitment to the leading edge. AU - Dow, L. AU - Kauffman, J. AU - Caddy, Jacinta. AU - Peterson, A. AU - Jane, Stephen. AU - Russell, Sarah. AU - Humbert, Patrick. PY - 2007. Y1 - 2007. M3 - Article. VL - 26. SP - 2272. EP - 2282. JO - Oncogene. JF - Oncogene. SN - 0950-9232. ER - ...
Hungwon Tchah is the author of this article in the Journal of Visualized Experiments: RNA Interference-based Investigation of the Function of Heat Shock Protein 27 during Corneal Epithelial Wound Healing
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Biochemical characterizationof thc pathway enzymes.(A) ThcE4/ThcD: FTMS spectra of chymotryptic digests ASSCDCSLY andGGCESCSYEGDEAE of ThcE4 modified by ThcD. E
... : A small purulent abscess was present under corneal epithelium in the substantia propria (or stroma of cornea) (H&E stain, original magnification x10 ...
Purpose: To describe a standardized, xenogenic-free protocol for the manufacture of limbal epithelial stem cell grafts and a no touch surgical technique for its standardized transplantation. Setting: Antwerp University Hospital, Antwerp, Belgium. Methods: The limbo-amnion composite graft is generated by cultivating limbal epithelial stem cells on a standardized (thermolysin treated and spongy layer removed) amniotic membrane, stretched within an interlockable amnion ring. The cells are cultured in CnT-20 medium with the addition of 1% human AB serum for a period of 2 weeks. Fibrin glue is applied to the surgically prepared recipients cornea and in one fluid motion, the composite graft within the amnion ring construct is transferred from culture and positioned onto the graft bed. The required size is cut out at the level of the limbus by means of a trephine and/or microsurgical scissors. Results: The lightweight, plastic interlockable ring offered stability to the graft during culture, ...
Limbal epithelial stem cells (LESCs) are tissue-specific stem cells responsible for renewing the corneal epithelium. Acute trauma or chronic disease affecting LESCs may disrupt corneal epithelial renewal, causing vision threatening and painful ocular surface disorders, collectively referred to as LESC deficiency (LESCD). These disorders cannot be treated with traditional corneal transplantation and therefore alternative cell sources for successful cell-based therapy are needed. LESCs derived from human pluripotent stem cells (hPSCs) are a prospective source for ocular surface reconstruction, yet critical evaluation of these cells is crucial before considering clinical applications. In order to quantitatively evaluate hPSC-derived LESCs, we compared protein expression in native human corneal cells to that in hPSC-derived LESCs using isobaric tag for relative and absolute quantitation (iTRAQ) technology. We identified 860 unique proteins present in all samples, including proteins involved in cell ...
Purpose: : Transplantation of ex vivo expanded limbal epithelial progenitor cells (LEPCs) on epithelially denuded amniotic membrane (dAM) in supplemented hormonal epithelial medium (SHEM) is an alternative solution for treating corneal blindness due to limbal stem cell (SC) deficiency. Because the phenotype of limbal niche cells (NCs) is preserved better in serum-free modified embryonic stem cell (ESC) medium (MESCM) than SHEM, we question whether the aforementioned expansion protocol can be further optimized by maintaining limbal NCs using MESCM. Methods: : Collagenase-isolated limbal clusters were cultured on dAM in SHEM or MESCM for 8 to 10 days. Epithelial outgrowth sheets removed by dispase were subjected to real-time quantitative polymerase chain reaction (qPCR) and immunostaining for expression of corneal epithelial markers (p63α, pax6, and K12) and NC markers (FLK-1, CD34, CD31, PDGFR-B, and α-SMA). A total of 1000 single cells were seeded on 6-well dish containing 3T3 feeder layers ...
Koizumi, N and Rigby, H and Fullwood, N J and Koizumi, K and Joussen, A M and Kociok, N (2004) Comparison of intact and denuded amniotic membrane as a substrate for human limbal epithelial cell culture. Investigative Ophthalmology and Visual Science, 45. U336-U336. ISSN 0146-0404 Full text not available from this repository ...
This study investigated DE-105 in patients with persistent corneal epithelial defect. The primary endpoint was restoration of corneal epithelial defect assessed
Recently, safety evaluation tests that do not involve animal experiments have been prosperously developing. However, the optimal evaluation materials and methods for assessing ocular irritancy have not been well investigated. In this study, we determined the optimal evaluation method for testing ocular irritation using a human cultured corneal epithelium model (corneal model). In order to assess adequate treatment conditions for the corneal model, we used cetylpyridinium chloride (CPC), which has been recognized as an irritant chemical by the Draize eye test. The irritancy elicited by multiple concentrations of CPC was evaluated by a cytotoxicity assay under nine treatment conditions and compared to the Draize score. The treatment conditions that included a 5-second exposure period followed by a 24-hour post-incubation period (hereafter called protocol "5-sec+24-h") showed a significant correlation between cytotoxicity and the Draize score. Furthermore, the dose-dependent cytotoxicity of six ...
A limbal biopsy taken from the contralateral good eye in cases of unilateral disease or from a living related or cadaveric donor in cases of bilateral disease. The limbal stem cells from the biopsy are cultivated until a sheet of cells measuring approximately 12mm in diameter is obtained. This is then ready for transplantation onto the diseased eye ...
In this study we evaluated the effects of Tualang honey (TH), an antioxidant-containing honey, on human corneal epithelial progenitor (HCEP) cells in culture. Cytotoxicity, gene expression, migration, and cellular resistance to oxidative stress were evaluated. Immunofluorescence staining revealed that HCEP cells were holoclonal and expressed epithelial stem cell marker p63 without corneal cytokeratin 3. Cell viability remained unchanged after cells were cultured with 0.004, 0.04, and 0.4% TH in the medium, but it was significantly reduced when the concentration was increased to 3.33%. Cell migration, tested using scratch migration assay, was significantly enhanced when cells were cultured with TH at 0.04% and 0.4%. We also found that TH has hydrogen peroxide (H2O2) scavenging ability, although a trace level of H2O2 was detected in the honey in its native form. Preconditioning HCEP cells with 0.4% TH for 48 h showed better survival following H2O2-induced oxidative stress at 50 µM than untreated ...
In this study we evaluated the effects of Tualang honey (TH), an antioxidant-containing honey, on human corneal epithelial progenitor (HCEP) cells in culture. Cytotoxicity, gene expression, migration, and cellular resistance to oxidative stress were evaluated. Immunofluorescence staining revealed that HCEP cells were holoclonal and expressed epithelial stem cell marker p63 without corneal cytokeratin 3. Cell viability remained unchanged after cells were cultured with 0.004, 0.04, and 0.4% TH in the medium, but it was significantly reduced when the concentration was increased to 3.33%. Cell migration, tested using scratch migration assay, was significantly enhanced when cells were cultured with TH at 0.04% and 0.4%. We also found that TH has hydrogen peroxide (H2O2) scavenging ability, although a trace level of H2O2 was detected in the honey in its native form. Preconditioning HCEP cells with 0.4% TH for 48 h showed better survival following H2O2-induced oxidative stress at 50 µM than untreated ...
It consists of several layers of cells. The cells of the deepest layer are columnar; then follow two or three layers of polyhedral cells, the majority of which are prickle cells similar to those found in the stratum mucosum of the cuticle. Lastly, there are three or four layers of squamous cells, with flattened nuclei. Epithelial ingrowth is a LASIK complication in which cells from the cornea surface layer (epithelial cells) begin to grow underneath the corneal flap. Epithelial ingrowth is a rarely occurring LASIK complication, appearing in less than one percent of LASIK procedures. This complication is not present in PRK or other non-flap vision correction procedures. Inner limiting membrane • Nerve fiber layer • Ganglion cell layer • Inner plexiform layer • Inner nuclear layer Outer plexiform layer • Outer nuclear layer . The corneal epithelium (epithelium corneæ anterior layer) is made up of epithelial tissue and covers the front of the cornea ...
Summary ArcScan technology was used to scan a cornea that had undergone radial keratotomy with inferior and superior trapezoidal keratotomies, resulting 27 years later in high irregular astigmatism (+6.50 -8.00 x 101) and severe loss of corrected distance visual acuity (CDVA) to 20/50. The epithelial thickness profile was highly irregular, masking a significant proportion of the true stromal […]. ...