Induced differentiation in vitro of rat ADSCs into photoceptors and RPE cells / 基础医学与临床

ABSTRACT

Objective To explore differentiation in vitro of rat adipose-derived stem cells into photoreceptor cells and RPE cells.Methods The ADSCs were cultured by adhering to the flask surface and purified by continual passaging.Surface antigens including CD45、CD90、CD49d、CD106 were indentified by flow cytometry.ADSCs were induced to differentiate by EGF,activin A,taurine,retinoic acid(RA) and extracted liquid of retina respectively.Meanwhile,ADSCs were induced by EGF+taurine,EGF+RA,taurine + RA,EGF+taurine+RA respectively.Immunofluorescence was used for detecting the expression of rhodopsin,CK and S-100,and flow cytometry was used for quantification.Results For primary culture,the phenotypes of ADSCs were CD45,CD90,CD49d and CD106,with a positive percentage of 1.6%,71.3%,7.8% and 3.5%,respectively.From passage 1 to 5,these phenotypes were CD45(0.8%～9.3%),CD90(84.7%～94.8%),CD49d(16.8%～31.0%)and CD106(8.3%～22.2%).There was a higher CD49d percentage than CD106 in all the passages.The induction efficacy of ADSCs was 17.5%～46.0% for rhodopsin,19.7%～79.3% for CK and 27.3%～50.7% for S-100.Conclusion It is suggested that ADSC has potential to differentiate into photoceptors and RPE cells as evidenced by thepresence of the specific markers of photoceptors(rhodopsin) and RPE markers(CK and S-100).