Duan Ping, Xu Haiwei, Zeng Yuxiao, Wang Yi, Yin Zheng Qin
Southwest Eye Hospital, Southwest Hospital, Third Military Medical University, Chongqing, China.
Cell Physiol Biochem. 2013;31(4-5):601-13. doi: 10.1159/000350080. Epub 2013 May 2.
There is an increasing interest in generating retinal pigment epithelial (RPE) cells from stem cells for therapy against degenerative eye diseases. Human bone marrow stromal cells (hBMSCs) can be induced to express retinal neuron-specific markers when co-cultured with retinal neurons, however, whether hBMSCs can differentiate into RPE-like cells in a co-culture system has not been clarified.
The induction of hBMSCs into RPE-like cells was performed by combining hBMSCs and pig RPE cells in a transwell system. The biomarkers of hBMSCs-derived RPE cells were determined by quantitative RT-PCR and immunofluorescence. The function of induced cells was assayed by ELISA for secretion of neurotrophic factors.
Intracellular pigment granules and many RPE markers existed in hBMSCs-derived RPE cells after co-culturing with pig RPE cells for 14 days. Typical RPE functions, such as phagocytosis of photoreceptor outer segments and secretion of the trophic factors, brain-derived neurotrophic factor (BDNF) and glia-derived neurotrophic factor (GDNF), were observed in these induced cells.
hBMSCs can be induced toward functional RPE cells simply by transwell-based co-culture with RPE cells.
利用干细胞生成视网膜色素上皮（RPE）细胞用于治疗退行性眼病的研究兴趣日益浓厚。人骨髓基质细胞（hBMSCs）与视网膜神经元共培养时可被诱导表达视网膜神经元特异性标志物，然而，hBMSCs在共培养系统中能否分化为RPE样细胞尚未明确。
通过在transwell系统中将hBMSCs与猪RPE细胞组合，诱导hBMSCs分化为RPE样细胞。采用定量RT-PCR和免疫荧光法检测hBMSCs来源的RPE细胞的生物标志物。通过ELISA检测诱导细胞分泌神经营养因子的功能。
hBMSCs与猪RPE细胞共培养14天后，其来源的RPE细胞内存在色素颗粒和多种RPE标志物。在这些诱导细胞中观察到了典型的RPE功能，如光感受器外段的吞噬作用以及神经营养因子、脑源性神经营养因子（BDNF）和胶质细胞源性神经营养因子（GDNF）的分泌。
通过基于transwell的与RPE细胞共培养，可将hBMSCs诱导为功能性RPE细胞。
