脐带源间充质干细胞与大鼠胰腺细胞共培养向胰岛样细胞分化及移植治疗1型糖尿病

背景：脐带Wharton’s Jelly中间充质干细胞可以向胰岛样细胞诱导分化。 目的：验证脐带源间充质干细胞与大鼠胰腺细胞共培养向胰岛样细胞诱导分化的可能性,并观察移植后对糖尿病大鼠血糖的影响。 方法：分离、诱导、传代脐带Wharton’s Jelly中间充质干细胞,再与大鼠胰腺细胞共培养,诱导成胰岛细胞团样组织。将大鼠分为3组,正常对照组不进行移植及造模;模型组仅制备糖尿病大鼠模型;实验组造模后将胰岛样细胞移植入糖尿病大鼠肾脏包膜。 结果与结论：脐带Wharton’s Jelly细胞培养中有细胞从组织块中爬出,第7天形态发生变化,贴壁细胞部分变成梭形。分离培养的细胞表达具有间充质干细胞表面特有标志CD44、CD29、CD105,不表达CD34、CD45、CD14。诱导第7,10天PDX-1及人胰岛素强染色;胰岛素及C-肽浓度较单纯培养组明显升高;PDX-1及人胰岛素mRNA诱导第7、10天较高表达。移植第1周大鼠尾尖血糖链脲佐菌素实验组明显低于模型组(P < 0.01),但明显高于正常照组(P < 0.01)。8周链脲佐菌素实验组肾脏被膜下发现胞核染棕色染色的Brdu阳性、胞浆棕色染色的胰岛素阳性细胞。结果表明,脐带Wharton’s Jelly中存在脐带源间充质干细胞,与大鼠胰腺细胞共培养可促进间充质干细胞向胰岛样细胞诱导分化,移植入糖尿病大鼠肾脏被膜下,可显著降低糖尿病大鼠血糖。

Islet-like cells derived from mesenchymal stem cells in Wharton's Jelly of the human umbilical cord co-cultured with rat pancreatic cells for transplantation to control

BACKGROUND:Mesenchymal stem cells in Wharton’s Jelly of the human umbilical cord can induce differentiation into islet-like cells. OBJECTIVE:To verify the possibility of human umbilical cord derived mesenchymal stem cells co-cultured with rat pancreatic cells differentiate into islet-like cells, and to observe the effects of transplantation of islet-like cells on blood glucose of diabetic rats. METHODS:Mesenchymal stem cells in Wharton’s Jelly of the human umbilical cord was separated, induced, passaged, and co-cultured with pancreatic cells to induce differentiation into islet-like clusters. Rats were divided into the normal control, model and experimental groups. Rats in the model group were prepared for diabetic models, and those in the experimental group were transplanted islet-like cells after model preparation. RESULTS AND CONCLUSION:There were cells crawled out of cultured Wharton’s Jelly of the human umbilical cord, and morphology of adhered cells turned into fusiform shape at 7 days. The isolated cells are characterized by expressing specific surface markers of mesenchymal stem cells, such as CD44, CD29, CD105, but not expressing CD34, CD45 or CD14. The cells were strongly stained by PDX-1 and human insulin at 7 and 10 days. Compared with the simple culture group, the expression of human insulin and concentration of C-peptide were obviously increased; PDX-1 and human insulin mRNA expressions were highly expressed at 7 and 10 days after induction. Compared with the model group, the streptozotocin test of rats in the experimental group was obvious decreased (P < 0.01), but extremely higher than that of the normal control group at 1 week after transplantation (P < 0.01). Brdu positive nuclei and insulin positive kytoplasms could be seen in the experimental group at 8 weeks after transplantation. The results demonstrated that, umbilical cord derived mesenchymal stem cells existed in Wharton’s Jelly. The co-cultured cells promote mesenchymal stem cells differentiating into islet-like cells, which can dramatically decrease blood glucose in diabetic rats.