Neurosphere-free Trans-differentiation of Rat Bone Marrow Stromal Stem Cells Into Retinal Cells and Retinal Pigment Epithelium

Background and Aim: Neurosphere-free (NF) trans-differentiation ofbone marrow stem cells (BMSCs) into retinal progenitor and retinal cellsin vitro could offer a unique opportunity for the study of regenerationof degenerative retinal diseases. In this respect, a simple and efficientprotocol for getting retinal cells from trans-differentiation of rat BMSCs inthe NF state is reported.Methods: BMSCs extracted from femur hooded pigment rats and passagedto P4 were exposed to single-step protocol, including neurosphere-free,which induced trans-differentiation into retinal pigment epithelium (RPE)and different retinal cells (RC). The cells immunostained with Rhodopsin,OTX2, NF200, GFAP, RPE65, VEGF, and CRALBP markers.Results: Cultured rat BMSCs at the fourth passage was shown to expressthe surface markers CD90 (95±2%), CD44 (91±2%), CD166 (98±2%),CD 106 (99±2%) and Fibronectin (96±2%), P < 0.05. The RC and RPEcells obtained from rat samples expressed the markers. 60 days afterdifferentiation, the previously rat BMSCs expressed rhodopsin (58±2%),OTX2 (24±2%), NF200 (25±2%), GFAP (28±2%), RPE65 (100±1%),CRALBP (96±1%), and VEGF (97±2%), P < 0.05 and were thussignificantly similar to the RC and RPE cells obtained from control ratsamples. Morphologically, differentiated cells appeared to have similarityto RC and epithelial and cytoplasmic pigment granules.Conclusion: Overall, these findings show a protocol, mimickingdevelopmental stages which make it possible to derive retinal and retinalpigment epithelium cells in vitro. This is a great finding for transplantationin retinal diseases.

کلیدواژه ها:

Neurosphere-Free ، Rat Bone Marrow Stem Cells ، Retinal Cells ، Retinal Pigment Epithelium ، Trans-differentiation