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目的:探讨人骨髓源干细胞向具有功能的胰岛素分泌细胞分化的可能性。方法:从人骨髓分离间充质干细胞。采用表皮生长因子、β-巯基乙醇和高糖培养基诱导其向胰岛素分泌细胞分化。经诱导后,用RT-PCR检测胰岛β细胞相关基因的表达,并采用免疫细胞化学染色检测胞浆胰岛素的表达。此外,诱导后细胞分泌的胰岛素定量及胰岛素释放实验将采用化学发光法进行检测。将经诱导后的细胞移植到糖尿病小鼠的右侧肾被膜下。在移植后16 d持续检测小鼠的血糖水平,最后对右侧肾脏进行免疫组化检测。结果:经诱导后,细胞能表达胰岛β细胞相关基因;免疫细胞化学染色也能检测到胞浆有胰岛素的表达;而且这些细胞能对糖刺激有所反应。细胞被移植到糖尿病小鼠的肾被膜下,能起降血糖作用。其肾脏的免疫组化显示:肾被膜下有胰岛素阳性细胞。结论:人骨髓源干细胞具有向胰岛素分泌细胞分化的潜能,这将为糖尿病细胞治疗提供丰富的细胞来源。
Objective: To explore the possibility of differentiation of human bone marrow-derived stem cells into functional insulin-secreting cells. Methods: Mesenchymal stem cells were isolated from human bone marrow. Epidermal growth factor, β-mercaptoethanol and high glucose medium were used to induce their differentiation into insulin-secreting cells. After induced, the expression of islet β cell related gene was detected by RT-PCR, and the expression of cytosolic insulin was detected by immunocytochemical staining. In addition, the amount of insulin secreted by the cells after induction and the insulin release assay will be determined by chemiluminescence. The induced cells were transplanted into the right kidney capsule of diabetic mice. At 16 days after transplantation, the blood glucose level in mice was measured continuously. Finally, the right kidney was examined by immunohistochemistry. Results: After induction, cells expressed islet β-cell related genes; immunocytochemical staining could detect the expression of insulin in cytoplasm; and these cells could respond to sugar stimulation. The cells are transplanted into the kidney capsule of diabetic mice, which can lower blood sugar. Immunohistochemistry of the kidneys showed that there were insulin-positive cells under the kidney capsule. CONCLUSION: Human bone marrow-derived stem cells have the potential to differentiate into insulin-secreting cells, which will provide a rich source of cells for the treatment of diabetic cells.