Ho JH, Tseng KC, Ma WH, Chen KH, Lee OK, Su Y (2008) Thymosin beta-4 upregulates anti-oxidative enzymes and protects human cornea epithelial cells against oxidative damage. Ho JH, Chuang CH, Ho CY, Shih YR, Lee OK, Su Y (2007) Internalization is essential for the antiapoptotic effects of exogenous thymosin beta-4 on human corneal epithelial cells.
Verstraelen J, Reichl S (2014) Multidrug resistance-associated protein (MRP1, 2, 4 and 5) expression in human corneal cell culture models and animal corneal tissue. īecker U, Ehrhardt C, Daum N, Baldes C, Schaefer UF, Ruprecht KW, Kim KJ, Lehr CM (2007) Expression of ABC-transporters in human corneal tissue and the transformed cell line, HCE-T. Kolln C, Reichl S (2012) mRNA expression of metabolic enzymes in human cornea, corneal cell lines, and hemicornea constructs. Yamasaki K, Kawasaki S, Young RD, Fukuoka H, Tanioka H, Nakatsukasa M, Quantock AJ, Kinoshita S (2009) Genomic aberrations and cellular heterogeneity in SV40-immortalized human corneal epithelial cells. Ronkko S, Vellonen KS, Jarvinen K, Toropainen E, Urtti A (2016) Human corneal cell culture models for drug toxicity studies. Juretic M, Jurisic Dukovski B, Krtalic I, Reichl S, Cetina-Cizmek B, Filipovic-Grcic J, Lovric J, Pepic I (2017) HCE-T cell-based permeability model: a well-maintained or a highly variable barrier phenotype? Eur J Pharm Sci 104:23–30. Toropainen E, Ranta V-P, Talvitie A, Suhonen P, Urtti A (2001) Culture model of human corneal epithelium for prediction of ocular drug absorption. īecker U, Ehrhardt C, Schneider M, Muys L, Gross D, Eschmann K, Schaefer UF, Lehr CM (2008) A comparative evaluation of corneal epithelial cell cultures for assessing ocular permeability. Reichl S (2008) Cell culture models of the human cornea - a comparative evaluation of their usefulness to determine ocular drug absorption in-vitro. Other cell models or differentiation protocols should be developed in the future to gain new tools for research on ocular surface diseases.Īraki-Sasaki K, Ohashi Y, Sasabe T, Hayashi K, Watanabe H, Tano Y, Handa H (1995) An SV40-immortalized human corneal epithelial cell line and its characterization. Primary LECs in the way cultured here are not an ideal system compared to differentiated epithelium in organ culture but should be preferred to HCE-T cells if corneal differentiation markers are investigated. Hence, this cell line is not suitable to study corneal differentiation processes. The HCE-T cell line is even less differentiated than LECs regarding the investigated markers and therefore might also lack the ability to express differentiation markers at protein level. PAX6 protein expression was hardly detected in HCE-T cells, and no difference could be seen between LECs and pCECs. DSG1 protein expression was only detected in pCECs. KRT3, KRT12, PAX6, ALDH1A1, ADH7, TP63, and CTSV mRNAs have shown increasing mRNA expression from HCE-T < HCE-T cultured in keratinocyte serum-free medium (KSFM) < LEC < to pCEC.KRT3 and KRT12 protein expressions were only slightly increased in LEC compared to HCE-T samples, and the strongest signals were seen in pCEC samples. KRT3, KRT12, DSG1, PAX6, ADH7, and ALDH1A1 mRNA expressions were higher in LECs and magnitudes higher in pCECs compared to HCE-T cells. Additionally, KRT3, KRT12, DSG1, and PAX6 protein levels were analyzed with Western blot. HCE-T cell line was purchased from RIKEN Institute RCB-2280.Expression levels of conjunctival- and corneal-specific keratin and adhesion markers (KRT3, KRT12, KRT13, KRT19, DSG1), stem cell and differentiation markers (PAX6, ABCG2, ADH7, TP63, ALDH1A1), and additional (unvalidated) putative differentiation and stem cell markers (CTSV, SPINK7, DKK1) were analyzed with qPCR.
Primary limbal epithelial cells (LECs) for cell culture and primary corneal epithelial cells (pCECs) as differentiated tissue samples were obtained from the limbus or central cornea region of corneal donors. This is necessary in order to decide whether HCE-T cells could be a tool to study the differentiation process and its regulatory networks in corneal epithelium. In this study, we want to compare the expression of differentiation markers in the HCE-T cell line to differentiated primary epithelial cells (pCECs) and primary limbal epithelial cell (LEC) culture. If these differentiation mechanisms are disturbed, it will lead to ocular surface disease.
The differentiation of (limbal) corneal epithelial into mature corneal epithelium coincides with the expression of established differentiation markers. Human corneal epithelial cell-transformed (HCE-T) cell line is used as a widely accepted barrier model for pharmacological investigations in the context of eye application.
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