Differentiation Of Rat Bone Marrow Mesenchymal Stem Cells Into Photoreceptors Like In Vitro

AIM: To find a technique yieds the highest percent of structurally well preserved cells in vitro. To isolate and culture rat mesenchymal stem cells (RMSCs) and induce them to differentiate into photoreceptor cells int vitro.METHODS: Photoreceptor cells from adult rat retinas were isolated and quantified using enzymatic dissociation without centrifugation.rMSCs were isolated and attached to the wall of culture dishes by their specific adherent ability.Then the cells were characterized by flow cytometry. rMSCs were induced by supernatant of retinal cell for 14 days in vitro.Then we used immunocytochemistry staining and RT-PCR to see whether they could express rhodopsin, NSE and GFAP.RESULTS: An enzymatic technique yielded the highest number of intact elongated photoreceptors right after dissociation. Many RMSCs,which were highly purified,were obtained after induction by supernatant of retinal cell,35.1%±6.2%RMSCs expressed rhodopsim,33.6%±4.0 % RMSCs expressed NSE, 9.6%±1.5% RMSCs expressed GFAP. In the controls,none RMSCs expressed rhodopsim,10.6%±5.0% RMSCs expressed NSE, 13.7%±3.0 % rMSCs expressed GFAP.CONCLUSION: The current study demonstrated the importance of proper isolation techniques to obtain the maximum amount of intact photoreceptors cells. The importance of proper isolation In vitro rMSCs are expanded, passaged and induced to differentiate into photoreceptor cell.