[In vitro transdifferentiation of adult human retinal pigment epithelium cells to neuro-like cells induced by blocking VEGF expression].
OBJECTIVE
To investigate the effects of blocking VEGF expression on the retinal pigment epithelium (RPE) in vitro.


METHODS
RPE was transfected with plasmid encoding anti-sense of VEGF by lipofectin. RPE clones with stable expression of anti-sense of VEGF were screened by neomycin (G418). The changes of RPE and the level of VEGF secreted by RPE were observed by immunofluorescence immunohistochemical staining, Elisa and inverted-microscope.


RESULTS
Transdifferentiation of cultured adult human retinal pigment epithelium cells to neurons was induced by blocking VEGF expression. The transdifferentiation was a direct phenotypic change from RPE cell to neurons without cell differentiation. The transdifferentiated neurons showed up-regulated expression of VEGF receptor 2 (Flk-1) and expressed several markers of retinal ganglion cell, including Thy1.1 and neurofilament.


CONCLUSIONS
Blocking of VEGF expression and up-regulation of Flk-1 can induce transdifferetiation of adult human retinal pigment epithelium cells into ganglion cells.
Wu
Ji-hong
JH
Central Experimental laboratory, The First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China.
Wang
Feng
F
Xu
Ping
P
Huang
Qian
Q
D000328:Adult; D002478:Cells, Cultured; D006801:Humans; D009474:Neurons / Q000166:cytology*; D016376:Oligonucleotides, Antisense; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism; D012160:Retina / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D016376:Oligonucleotides, Antisense; D042461:Vascular Endothelial Growth Factor A