Abstract
Cryopreserved retinal pigment epithelial (RPE) cells at -80°C can be used for transplantation studies. Long-term viability at -196°C has been used for many other cell lines. The origin and purity of human RPE cell lines were assessed by immunocytochemistry and reverse-transcriptase polymerase chain reaction (RT-PCR) using RPE cell markers [cytokeratin, cellular retinaldehyde binding protein (CRALBP), tyrosinase, tyrosinase-related proteins I and II (TRP-I and II), Na,K-ATPase α1 and β2]. Cultured human RPE cell lines were cryopreserved at -80°C and -196°C. After cryopreservation, human RPE cells were re-cultured on laminin-coated polystyrene dishes and laminin-coated collagen sheets. Differentiation was evaluated by the expression level of RPE marker genes using RT-PCR. It was found that cryopreserved human RPE cells on laminin-coated collagen sheets disclosed relatively strong re-expression of their marker genes if compared to human RPE cells cultured on laminin-coated polystyrene dishes. Cryopreservation of RPE cells at both temperatures, thawing and subsequent culturing on laminin-coated collagen sheets can enhance the feasibility of using cryopreserved RPE cells for transplantation.
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© 1999 Kluwer Academic / Plenum Publishers
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Durlu, Y.K. et al. (1999). The Retinal Pigment Epithelial Cell Differentiation and Cell Marker Expression Following Cryopreservation at -80°C and under Liquid Nitrogen at -196°C. In: Hollyfield, J.G., Anderson, R.E., LaVail, M.M. (eds) Retinal Degenerative Diseases and Experimental Therapy. Springer, Boston, MA. https://doi.org/10.1007/978-0-585-33172-0_53
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DOI: https://doi.org/10.1007/978-0-585-33172-0_53
Publisher Name: Springer, Boston, MA
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