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The Retinal Pigment Epithelial Cell Differentiation and Cell Marker Expression Following Cryopreservation at -80°C and under Liquid Nitrogen at -196°C

  • Y. K. Durlu
  • S. -I. Ishiguro
  • K. Akaishi
  • T. Abe
  • Y. Chida
  • S. Shibahara
  • M. Tamai

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.

Keywords

Retinal Pigment Epithelial Retinal Pigment Epithelial Cell Human Retinal Pigment Epithelial Cell Human Retinal Pigment Epithelial Cell Marker Expression 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Kluwer Academic / Plenum Publishers 1999

Authors and Affiliations

  • Y. K. Durlu
    • 1
  • S. -I. Ishiguro
    • 1
  • K. Akaishi
    • 1
  • T. Abe
    • 1
  • Y. Chida
    • 1
  • S. Shibahara
    • 2
  • M. Tamai
    • 1
  1. 1.Department of OphthalmologyTohoku University School of MedicineSendai, MiyagiJapan
  2. 2.Department of Molecular BiologyTohoku University School of MedicineSendai, MiyagiJapan

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