Transfection of basic fibroblast growth factor (bFGF) gene or bFGF antisense fene into human fetinal pigment epithelial cells View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1999-07

AUTHORS

N. Ogata, Mikio Nishizawa, Akira Ando, Masanobu Uyama

ABSTRACT

BACKGROUND: Transplantation of RPE cells offers a potential of restoring retinal pigment epithelium (RPE) function and has been shown to be effective in the dystrophic RCS rat model. Recently, RPE transplantation was attempted in patients with age-related macular degeneration. Basic fibroblast growth factor (bFGF) plays important roles in maintaining normal retinal function. The purpose of this study was to introduce bFGF sense or antisense cDNA into human RPE cells to alter the expression of bFGF. METHODS: Human bFGF sense cDNA or antisense cDNA was inserted into the pBK-CMV vector. For stable gene expression, we introduced the plasmids into RPE cells using the electroporation method. Following electroporation, transfected RPE cells were cultured and resistant cells were selected in the presence of antibiotic G418. We analyzed the expression of the transfected genes in the cloned RPE cells by polymerase chain reaction (PCR) and by reverse transcription (RT)-PCR. RESULTS: Cloned RPE cells in which the bFGF sense or antisense cDNA had been efficiently transfected were established. PCR and RT-PCR analysis demonstrated not only the presence but also the expression of bFGF sense or antisense cDNA in the transfected RPE cells. CONCLUSIONS: Human bFGF sense cDNA or antisense cDNA can be efficiently introduced into cultured RPE cells by the electroporation method. The successful expression of the genes into RPE cells demonstrated that this technique can be used to regulate bFGF expression and thus increase the scope of RPE transplantation for the treatment of retinal diseases. More... »

PAGES

678-684

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s004170050296

DOI

http://dx.doi.org/10.1007/s004170050296

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1015249324

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/10459618


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