Guoji Yanke Zazhi (Mar 2014)
<i>In vitro</i> differentiation of MSCs into retina-like cells by the supernatant fluid of light-injured neurosensory retina
Abstract
AIM: To explore the possibility of inducing rat mesenchymal stem cells(MSCs)into retina-like cells by the supernatant fluid of light-injured neurosensory retina in vitro.METHODS: MSCs were isolated and attached to the wall of culture dishes by their specific adherent ability. Then the cells were characterized by flow cytometry. The neurosensory retina was isolated from retina of SD rat and it was tested by hematoxylin-eosin(HE)staining. The pathological changes of light-injured neurosensory retina was observed under transmission electron microscope. Three kinds of supernatant fluid of light-injured neurosensory retina of SD rats were prepared. The third passage of MSCs were cultured with these mixed medium for 7-8d, we used RT-PCR to see whether they could express rhodopsin, neuron-specific enolase(NSE), and glial fibrillary acidic protein(GFAP), and positive cells were counted and analyzed.RESULTS: HE staining showed the retinal sheets included full-thickness neural retina. Neurosensory retina developed ultrastructural destructions by light injury. RT-PCR showed that the medium of mixed I expressed higher positive rate of rhodopsin(0.3915±0.00644), NSE(0.2019±0.00682), GFAP(0.1972±0.00211)than the medium of mixed Ⅱ rhodopsin(0.0983±0.00319), NSE(0.1048±0.00323), GFAP(0.1040±0.00254)and medium of mixed Ⅲ rhodopsin(0.0044±0.00126), NSE(0.0498±0.00149), GFAP(0.0467±0.00333). The difference of intergroup has statistical significance.CONCLUTION:The supernatant fluid of light-injured neurosensory retina of SD rats can induce MSCs to differentiate into retina-like cells and provide new insights of stem cell therapy for retinopathy.
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