AIM: To explore the possibility of marrow mesenchymal stem cells (MSC) in vitro differentiating into functional islet-like cells and to test the diabetes therapeutic potency of Islet-like cells.METHODS: Rat MSCs were isolated from Wistar rats and cultured. Passaged MSCs were induced to differentiate into islet-like cells under following conditions: pre-induction with L-DMEM including 10 mmol/L nicotinamide + 1 mmol/L β –mercaptoethanol + 200 mL/L fetal calf serum (FSC) for 24 h, followed by induction with serum free H-DMEM solution including 10 mmol/L nicotinamide + 1 mmol/L,β -mercaptoethanol for 10 h. Differentiated cells were observed under inverse microscopy, insulin and nestin expressed in differentiated cells were detected with immunocytochemistry. Insulin excreted from differentiated cells was tested with radioimmunoassay. Rat diabetic models were made to test in vitro function of differentiated MSCs.RESULTS: Typical islet –like clustered cells were observed. Insulin mRNA and protein expressions were positive in differentiated cells, and nestin could be detected in pre-differentiated cells. Insulin excreted from differentiated MSCs (446.93 ± 102.28 IU/L) was much higher than that from pre-differentiated MSCs (2.45 ± 0.81 IU/L (P < 0.01). Injected differentiated MSCs cells could down-regulate glucose level in diabetic rats.CONCLUSION: Islet-like functional cells can be differentiated from marrow mesenchymal stem cells, which may be a new procedure for clinical diabetes stem -cell therapy, these cells can control blood glucose level in diabetic rats. MSCs may play an important role in diabetes therapy by islet differentiation and transplantation.
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机译：目的：探讨骨髓间充质干细胞体外分化为功能性胰岛样细胞的可能性，并检验胰岛样细胞对糖尿病的治疗作用。方法：从Wistar大鼠中分离培养大鼠MSC。在以下条件下诱导传代的MSC分化为胰岛样细胞：用包含10 mmol / L烟酰胺+ 1 mmol / Lβ-巯基乙醇+ 200 mL / L胎牛血清（FSC）的L-DMEM预诱导24 h然后用含10 mmol / L烟酰胺+ 1 mmol / L，β-巯基乙醇的无血清H-DMEM溶液诱导10 h。倒置显微镜下观察到分化的细胞，用免疫细胞化学法检测分化细胞中胰岛素和巢蛋白的表达。用放射免疫测定法测试从分化细胞中分泌的胰岛素。建立大鼠糖尿病模型，检测分化的MSCs的体外功能。结果：观察到典型的胰岛样簇细胞。分化细胞中胰岛素mRNA和蛋白质表达阳性，预分化细胞中可检测到巢蛋白。分化后的MSCs分泌的胰岛素（446.93±102.28 IU / L）比分化前的MSCs分泌的胰岛素（2.45±0.81 IU / L）高（P <0.01），注射的分化后的MSCs可以下调糖尿病大鼠的血糖水平。结论：可将胰岛样功能细胞与骨髓间充质干细胞区分开来，这可能是临床治疗糖尿病干细胞的新方法，这些细胞可控制糖尿病大鼠的血糖水平，MSCs可能在糖尿病治疗中起重要作用通过胰岛分化和移植。
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