大鼠的两种干细胞向光感受器细胞和视网膜色素上皮细胞分化的实验研究

目的研究大鼠的骨髓间充质干细胞（ MSCs ）和脂肪干细胞（ ADSCs ）体外向光感受器细胞和RPE细胞分化的可能性，为临床治疗视网膜病变提供物质基础。方法经贴壁筛选法连续传代纯化细胞，流式细胞仪检测CD45、CD90、CD49d、CD106鉴定MSCs和ADSCs。用视网膜细胞提取液、EGF、牛磺酸和视黄酸分别诱导MSCs和ADSCs；流式细胞仪定量检测细胞的视紫红质、CK和S-100的表达，计算诱导效应。结果传代纯化MSCs和ADSCs，传1代至传5代，MSCs中CD45阳性率是1．8％～7．5％，CD90是83．8％～95．7％，CD49d是6．6％～24．8％，CD106是15．7％～32．2％；ADSCs中CD45阳性率是0．8％～9．3％，CD90是84．7％～94．8％，CD49d是16．8％～31．0％，CD106是8．3％～22．2％。 MSCs和ADSCs 均为CD45阴性和CD90阳性。各代MSCs和ADSCs比较，MSCs低表达CD49d，高表达CD106；而ADSCs高表达CD49d，低表达CD106。不同的诱导方法，MSCs的视紫红质的诱导效应是55．9％～70．0％，CK的诱导效应是49．5％～72．0％，S-100的诱导效应是25．4％～34．0％；ADSCs的视紫红质的诱导效应是19．6％～31．1％，CK的诱导效应是31．0％～79．3％，S-100的诱导效应是27．3％～50．7％。结论 MSCs和ADSCs均具有向光感受器细胞和RPE细胞分化的潜能，可能成为干细胞移植治疗的来源。

Differentiation of rat two-derived stem cells into photoceptors and RPE cells

Objective To explore method to isolate and culture bone marrow mesenchymal stem cells (MSCs) and adipose-derived stem cells(ADSCs), and in-vitro to induce them into photoceptors and RPE cells .To provide material ba-sis for the clinical treatment of retinal diseases .Methods The MSCs and ADSCs were cultured by adhering to the flask surface and purified by continual passage .Surface antigens including CD45、CD90、CD49d、CD106 were indentified by flow cytometry.MSCs and ADSCs were induced to differentiate by extracted liquid of retina ,EGF, taurine and retinoic acid (RA)respectively.Flow cytometry was used for quantifying the expression of rhodopsin , CK and S-100 and the induction efficacy.Results Purified from passage 1 to 5, MSCs phenotypes were: CD45(1.8%~7.5%), CD90(83.8%~95. 7%), CD49d(6.6%~24.8%)and CD106(15.7%~32.2%); ADSCs phenotypes were: CD45(0.8%~9.3%), CD90(84.7%~94.8%), CD49d(16.8%~31.0%)and CD106(8.3%~22.2%).In all the passages, there was a lower CD49d percentage than CD106 in MSCs and a higher CD49d percentage than CD106 in ADSCs contrarily.The induc-tion efficacy of MSCs was 55.9%~70.0%for rhodopsin, 49.5%~72.0%for CK and 25.4%~34.0%for S-100 and of ADSCs was 19.6%~31.1%for rhodopsin, 31.0%~79.3%for CK and 27.3%~50.7%for S-100.Conclusion It is suggested that MSCs and ADSC have potential to differentiate into photoceptors and RPE cells .Both of them may represent sources of stem cells transplantation for retinopathy.