[Isolation and identification of human embryonic retinal stem cells].
OBJECTIVE
To isolate and identify human retinal stem cells (HRCs) from human embryonic retina.


METHODS
HRCs were isolated from neural retinas at the 16th-20th week of gestation and cultured in either serum-free or serum-containing media. The expression of nestin antigen and retinal terminal cell markers were conformed by immunofluorescence labeling technique. RT-PCR analysis was used to identify the difference of nestin gene expression in HRCs and differentiated cells.


RESULTS
HRCs divided into multiple generations (up to passage 8) and expressed nestin. Other characteristics of the isolated cells were to differentiate into glial fibrillary acidic protein-positive glial cells (Müller cells) and retina-specific neurons expressing rhodopsin, syntaxin, PKCalpha, and Thy1. These were markers for rod receptors, amacrines, bipolar cells, and ganglion cells respectively. When cultured with serum, the expression of nestin was significantly downregulated in HRCs.


CONCLUSIONS
Human neural retina at the 16th-20th week of gestation harbors neural stem cells that are capable of selfrenewal and multilineage differentiation.
Yu
Hai-yan
HY
Department of Ophthalmology, Third Hospital, Beijing Medical University, Beijing 100083, China.
Shen
Li
L
Wang
Wei
W
D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin