摘要
Objective: To investigate the immunocytochemical characteristics of cultured human retinal pigment epithelial cells (RPEC). Methods: Indirect immunofluorescence mehtods were applied to study the expression of keratin, vimentin, actin, Ⅷ factor and glial fibrillary acidic protein (GFAP), and HAM45 in the 1st-passage and the 3rd- to 6th-passages of RPEC. Results: subcultured human RPEC demonstrated positive staining for keratin and vimentin protein; keratin reactivity was a constant feature of all human RPEC.The first-passage RPEC did not stain with abtibodies to vimentin, or stained very weakly. After passaged for 3 to 6 times, however, RPEC demonstrated intensively positive staining for vimentin. BPEC expressed generally negative staining for actin, Ⅷ factor, GFAP,and HAM45. Conclusion: Cultured RPECs always express keratin other than proteins of actin, Ⅷ factor, GFAP and HAM45. Protein vimentin expression intensity of RPEC increases with more passages. This indicates alternative phenotype of RPEC due to cell cultural conditions.
Objective: To investigate the immunocytochemical characteristics of cultured human retinal pigment epithelial cells (RPEC). Methods: Indirect immunofluorescence mehtods were applied to study the expression of keratin, vimentin, actin, Ⅷ factor and glial fibrillary acidic protein (GFAP), and HAM45 in the 1st-passage and the 3rd- to 6th-passages of RPEC. Results: subcultured human RPEC demonstrated positive staining for keratin and vimentin protein; keratin reactivity was a constant feature of all human RPEC.The first-passage RPEC did not stain with abtibodies to vimentin, or stained very weakly. After passaged for 3 to 6 times, however, RPEC demonstrated intensively positive staining for vimentin. BPEC expressed generally negative staining for actin, Ⅷ factor, GFAP,and HAM45. Conclusion: Cultured RPECs always express keratin other than proteins of actin, Ⅷ factor, GFAP and HAM45. Protein vimentin expression intensity of RPEC increases with more passages. This indicates alternative phenotype of RPEC due to cell cultural conditions.
基金
National Natural Science Foundation of China. No.39700154