The Retinal Pigment Epithelial Cell Differentiation and Cell Marker Expression Following Cryopreservation at -80°C and under Liquid Nitrogen at -196°C
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.
KeywordsRetinal Pigment Epithelial Retinal Pigment Epithelial Cell Human Retinal Pigment Epithelial Cell Human Retinal Pigment Epithelial Cell Marker Expression
Unable to display preview. Download preview PDF.
- 4.Y.K. Durlu and M. Tamai, 1995, In vitro expression of epidermal growth factor receptor by human retinal pigment epithelium cells, in: Degenerative Diseases of the Retina (R.E. Anderson, M.M. La Vail, J.G. Hollyfield, eds.), pp. 69–76, Plenum Press, New York.Google Scholar
- 7.S.H. Sarks and J.P. Sarks, 1989, Age-related macular degeneration: atrophic form, in: Retina (S.J. Ryan, ed.), Volume 2, pp. 149–173, The C. V. Mosby Co., St. Louis.Google Scholar