功能性胰岛素分泌细胞化学诱导方案的研究

Research on the chemical induction scheme for functional insulin producing cell

  • 摘要:
    目的  探讨脂肪间充质干细胞(ADMSC)向胰岛素分泌细胞(IPC)分化的有效诱导方案。
    方法  使用不同方案的小分子化合物诱导分化ADMSC,通过流式分析细胞纯度,镜下观察细胞形态学变化,苏木素-伊红和免疫组织化学染色检测细胞质量、性能及胰岛素相关指标,双硫腙(DTZ)与二乙酰荧光素/碘化丙啶染色检测细胞成熟度及活性,分析ADMSC向IPC分化诱导效果。
    结果  ADMSC纯度可达99%以上,方案Ⅰ、Ⅱ、Ⅲ的成球性良好;细胞诱导质量及胰腺十二脂肠同源盒1(PDX1)、肌腱膜纤维肉瘤癌基因同源物A(MAFA)、胰岛素和C肽表达效果均为方案Ⅰ优于其它方案;DTZ染色深浅可能与IPC成熟度有关,其中方案Ⅰ的凋亡细胞数量明显较方案Ⅱ、Ⅲ少。
    结论  诱导方案Ⅰ能提高ADMSC向IPC的分化效率,为未来临床IPC移植应用奠定一定基础。

     

    Abstract:
    Objective  To explore the effective induction scheme for differentiation of adipose-derived mesenchymal stem cell (ADMSC) to insulin producing cell (IPC).
    Methods  Different schemes of small molecule compound were used to induce the differentiation of ADMSC. The purity of cells was analyzed by flow cytometry and the morphological changes of cells were observed under the microscope. The quality, performance and insulin related indicators of cells were detected by hematoxylin-eosin and immunohistochemical staining. The maturity and activity of cells were detected by dithizone (DTZ) and diacetylfluorescein/propidium iodide staining. The induction effect of ADMSC differentiated into IPC was analyzed.
    Results  The purity of ADMSC reached more than 99%, and the sphere forming properties of schemes Ⅰ, Ⅱ, and Ⅲ were good. Cell induction mass, the expression effects of pancreatic and duodenal homeobox 1 (PDX1), musculoaponeurotic fibrosarcoma oncogene homolog A (MAFA) and insulin and C peptide of schemes Ⅰ were both better than those of other schemes. The DTZ staining depth may be related to IPC maturity, among which the number of apoptotic cells in scheme Ⅰ was significantly less than that of scheme Ⅱ and Ⅲ.
    Conclusions  Induction scheme Ⅰ may improve the differentiation efficiency of ADMSC to IPC and lay a certain foundation for future clinical IPC transplantation applications.

     

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