In Vitro Differentiation Of Rat Mesenchymal Stem Cells Into Retina-like Cells By The Supernatant Fluid Of Light-injured Neurosensory Retina

Objective:To explore the possibility of inducing SD rat mesenchymal stem cells (MSCs) into retina-like cells by the supernatant fluid of light-injured neurosensory retina in vitro.Methods:1,MSCs were isolated and attached to the wall of culture dishes by their specific adherent ability,Then the cells were characterized by flow cytometry. 2,The neurosensory retina was isolated from retina of SD rat and it was tested by hematoxylin-eosin (HE) staining. 3,The neurosensory retina was induced light injury by exposed to light with intensity of (1950±200)Lux for 45minutes, The pathological changes of light-injured neurosensory retina was observed under transmission electron microscope.4,The supernatant fluid of light-injured neurosensory retina of SD rats were collected, sterilized and mixed with 10%FBS-LGDMEM medium according to 2:3 proportion, The third passage of MSCs were cultured with these mixed medium for 7~8 days, 5,The differentiated retina-like cells were identified with inverted mircroscopy. 6,we used immunocytochemistry staining and RT-PCR to see whether they could express Rhodopsin , NSE(neuron-specific enolase),and GFAP(glial fibrillary acidic protein),and positive cells were counted and analysed Results:1,Many MSCs which were highly purified were obtained. 2,Hematoxylin-eosin staining showed the retinal sheets included full-thickness neural retina.3,Neurosensory retina developed ultrastructural destructions by light injury.4,The cell was observed by inverted mircroscopy after7~8days,the medium of mixedⅠcan induced MSCs appear ecphyma.Part of them was moving and forming synapse contect. 5,Immunocytochemistry staining showed that the medium of mixedⅠexpressed higher positive rate of Rhodopsin (37.97±7.84)%,GFAP(20.73±5.25)%,NSE(21.59±2.98)% than the medium of mixedⅡand medium of mixedⅢ,.The difference of intergroup has statistical significance.The results from RT-PCR were similar to immunocytochemistry staining.Conclution:1,The supernatant fluid of light-injured neurosensory retina of SD rats can induce MSCs to differentiate into retina-like cells.2,These MSCs-converted retina-like cells showed some of the characteristics of retina cells.