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Haibin Tian, Peng Li, Li Wang, Chunpin Lian, Weiwei Wang, Caixia Jin, Lixia Lu, Weiye Li, Guo-Tong Xu; Differentiaiton of human cord mesenchymal stem cells into retinal cells in porcine retinal pigment epithelial cell conditioned media. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2221.
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© ARVO (1962-2015); The Authors (2016-present)
Human cord mesenchymal stem cells (hCMSCs) have multipotential differentiation capacity. Transplantation of hCMSC derived retinal cells provides promise for prevention or cure of retinal degeneration (RD). The functions of porcine retinal pigment epithelial cell (RPE) conditioned medium on differentiation of hCMSCs into retinal cells were investigated.
hCMSCs were cultured in DMEM/F12 medium plus 10% FBS and specific markers for mesenchymal stem cells on hCMSCs were confirmed by FACS. Porcine RPE conditioned media were obtained by culturing porcine RPE cells in DMDM/F12 plus N2 (RPE-N2 conditioned medium) or 2%FBS (RPE-FBS conditioned medium) for 24h. For differentiation, hCMSCs were cultured in RPE-N2 conditioned medium or RPE-FBS conditioned medium for two weeks. hCMSC derived RPE-like cells and photoreceptor-like cells were confirmed by detecting the expressions of specific markers via Q-PCR, Westernblot and immunostaining. RPE-like cells were further confirmed by phagocytosis of FITC-labeled photoreceptor outer segments (POS).
hCMSCs were positive for CD105, CD90, CD73, CD44, CD29 and negative for CD34, CD45, hCMSCs were positive for CD105, CD90, CD73, CD44, CD29 and negative for CD34, CD45, MHCII. When hCMSCs were cultured in porcine RPE-N2 conditioned medium, neural differentiation pathway was triggered, which was confirmed by upregulated expressions of nestin, MAP-2 and photoreceptor specific marker Rho. However, RPE-FBS conditioned medium promoted hCMSCs to differentiate into RPE65+Mitf+CK8/18+ RPE-like cells. Pigments were not observed in these differentiated cells. Compared to hCMSCs, RPE-like cells can phagocyte much more POS.
RPE conditioned media can promote hCMSCs to differentiate into RPE-like or photoreceptor-like cells under different conditions, which will provide enough cells for clinical treatment of patients with RD.
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