论文部分内容阅读
目的研究人脐带间充质干细胞(human umbilical cord Wharton s jelly-derived mesenchymal stem cells,HuMSCs)向胰岛素分泌细胞分化的潜能,为胰岛细胞移植治疗Ⅰ型糖尿病提供新的细胞来源。方法体外分离培养HuMSCs,在胰岛细胞培养条件下经药物(尼克酰胺、β-巯基乙醇和高糖)定向诱导其分化;倒置相差显微镜下观察诱导后细胞形态;免疫组化、RT-PCR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;放射免疫法检测胰岛素含量。结果诱导后HuMSCs由长梭形逐渐变为多角形、类圆形,部分聚集成团;免疫细胞化学染色显示,HuMSCs经诱导后表达人胰岛素和胰高血糖素抗原;RT-PCR检测显示诱导后HuMSCs表达了人胰岛素基因和PDX-1基因;双硫腙染色呈棕红色。结论HuMSCs在体外胰岛细胞培养条件诱导下,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。
Objective To study the potential of human umbilical cord Wharton ’s jelly-derived mesenchymal stem cells (HuMSCs) to differentiate into insulin-secreting cells and to provide a new source of cells for islet cell transplantation in the treatment of type 1 diabetes. Methods HuMSCs were isolated and cultured in vitro. The cells were induced to differentiate under the condition of islet cell culture (Nicotinamide, β-mercaptoethanol and high glucose). The morphological changes of the cells were observed under inverted phase contrast microscope. Immunohistochemistry and RT- The cells were identified by islet β-cell labeling. Dithizone staining was used to identify the expression of zinc ion and the radioimmunoassay was used to detect the content of insulin. Results After induction, HuMSCs gradually transformed from long fusiform to polygonal, round and partially aggregated into clusters. Immunocytochemical staining showed that HuMSCs expressed human insulin and glucagon antigen after induction; RT-PCR showed that after induction HuMSCs express the human insulin gene and the PDX-1 gene; dithizone staining is reddish brown. Conclusion HuMSCs have the potential to differentiate into insulin-secreting cells under the condition of islet cell culture in vitro, which may provide a new therapeutic approach for type I diabetes.