体外诱导胎儿骨髓间充质干细胞向胰岛β样分泌细胞的分化

背景：体外诱导人骨髓间充质干细胞定向分化为胰岛样细胞,目前尚无成熟的鉴定方案。 目的：探讨胎儿骨髓间充质干细胞在适当的条件下体外分化为胰岛样分泌细胞的可能性。   方法：将胎儿骨髓间充质干细胞与胎儿胰岛素细胞分别接种于Transwell 双层细胞培养板上下层共培养。对照组中的骨髓间充质干细胞仅用胰岛细胞培养基培养。倒置相差显微镜观察胎儿骨髓间充质干细胞、胎儿胰岛细胞的形态,放射免疫分析法检测共培养刺激下胰岛素的分泌情况。 结果与结论：未经诱导的骨髓间充质干细胞呈长梭形贴壁生长,与胰岛细胞共培养的骨髓间充质干细胞逐渐变圆,并聚集成团,共培养9 d后骨髓间充质干细胞经RAI检测分泌大量胰岛素,DTZ染色为阳性,细胞免疫化学检测阳性。而对照组无胰岛素释放。提示在适当的培养条件下,胎儿骨髓间充质干细胞具有向胰岛素分泌细胞分化的能力。

In vitro differentiation of fetal bone marrow mesenchymal stem cells into islet beta like cells

BACKGROUND:There is no a mature program for identifying islet like cells differentiated from human bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE:To investigate the possibility of human fetal BMSCs differentiating into insulin-secreting cells in appropriate condition. METHODS:The fetal BMSCs were co-cultured with fetal insulin cells on Transwell double-layer culture plate. In the control group, the BMSCs were cultured in culture medium containing islet cells. Morphological changes in fetal BMSCs and islet cells were analyzed under a phase contrast microscopy. The morphological changes and insulin secretions of BMSCs under the stimulation of fetal islet cells were detected by radioimmunoassay. RESULTS AND CONCLUSION:The fetal BMSCs were isolated and purified, identical cell morphology liked plants spindle-like arrangement. Fetal islet cells were confirmed by panccreatic β endocrine cells under electron microscopy. The undifferentiated BMSCs exhibited were adherent and long spindle-shaped cells. After induction, cells gradually became round and formed clusters. Cells secreted abundant insulin and formed islet-like cell clusters that exhibited positive dithizone staining and immunocytochemical staining at 9 days. No insulin was detected in the control group. It is indicated that fetal BMSCs have the potentiality to differentiate into insulin-producing cells in appropriate condition．