王英+罗浩虹+高洪泉+缪智辉+赵亚清

【摘要】目的研究人脐带血间充质干细胞向胰岛素分泌细胞分化的潜能及其移植后对糖尿病大鼠的治疗效果。方法体外分离培养HUCB-MSCs, 在胰岛细胞培养条件下经药物定向诱导其分化；免疫组化对诱导细胞进行胰岛β细胞标记鉴定；双硫腙染色鉴定锌离子表达及检测胰岛样细胞的移植效果。结果HUCB-MSCs经诱导后, 免疫细胞化学染色显示表达人胰岛素；双硫腙染色呈棕红色；移植后2周, 胰岛样细胞组血糖浓度明显降低。结论HUCB-MSCs在体外诱导培养条件下, 具有向胰岛素分泌细胞分化的潜能, 这种细胞可能为Ⅰ型糖尿病提供一条新的治疗途径。

【关键词】人脐带血；间充质干细胞；定向诱导；糖尿病

Inducing human umbilical cord blood mesenchymal stem cells into insulin secreting cells in vitro culture and its effects on diabetes rats WANG Ying, LUO Hao-hong, GAO Hong-quan, et al. Medical College of Xiamen, Xiamen 361008, China

【Abstract】ObjectiveTo explore the possibility of inducing the differentiation of human umbilical cord blood mesenchymal stem cells(HUCB-MSCs) into insulin secreting cells in vitro and the therapeutic efficacy on diabetic rats following transplantation. Methods We isolated and cultured HUCB-MSCs, and induced HUCB-MSCs to differentiate into insulin secreting cells in the islet cell culture condition. HUCB-MSCs of induction were detected by immunocytochemical Methods. Zinchydronium was detected by Dithizon staining. Transplantation efficiency of islet-like cells was detected. Results Under induction, immunocytochemical examination showed that the expression of human insulin was positive. Dithizon stain showed that the cytoplasm of HUCB-MSCs was stained in brownish red color after induction. At 2 weeks following transplantation, blood glucose concentration in the islet-like cell group significantly reduced. Conclusion HUCB-MSCs can be differentiated into insulin secreting cells in vitro. HUCB-MSCs have the potential to become an excellent candidate in β cell replacement therapy of typeⅠdiabetes.

【Key words】Human umbilical cord blood; Mesenchymal stem cells; Induction; Diabetes rats2000年, Erices等［1］报道人脐血中含有丰富的造血干细胞和间充质干细胞(mesenchymal stem cell/ mesenchymal progenitor cell, MSC/MPC)。脐带血间充质干细胞(umbilical cord blood mesenchymal stem cells, UCB-MSCs)为中胚层发育的早期细胞, 具有多向分化的潜在能力, 其形态和生物学特点与骨髓来源 MSCs 相似［2］。UCB-MSCs具备自我更新和增殖的能力, 并具备多向分化潜能特性。在一定条件下进行体外培养和诱导分化后具有向神经细胞(神经元和神经胶质细胞)、成骨细胞、软骨细胞、脂肪细胞和心肌细胞等方向分化功能, 可作为细胞治疗的种子细胞和基因治疗的载体细胞［3-5］。脐血又具有淋巴细胞抗原性弱、功能相对不成熟、移植后发生移植物抗宿主病的风险较低等特点, 与其他类型的干细胞比较, 取材方便, 扩增数大, 来源广泛, 不受伦理和道德的限制, 因此脐血干细胞将以其自身的特性和优势占据重要的位置, 具有重大的理论意义和潜在的临床应用价值［6］。……