人脐血间充质干细胞的冻存复苏及向肝细胞分化的研究

发布时间：2018-04-14 11:16

本文选题：脐血 + 间充质干细胞　；参考：《四川大学》2006年硕士论文

【摘要】：目的：研究冻存复苏对人脐血间充质干细胞(UCBMSCs)生长状况的影响；研究复苏后的UCBMSCs在肝细胞生长因子(HGF)和成纤维细胞生长因子-4(FGF-4)的联合诱导下，向肝细胞分化的潜能，以及类肝细胞对UCBMSCs是否具有定向诱导的作用。方法： 1．用含10％二甲基亚砜(DMSO)的细胞培养基将第6代UCBMSCs冻存于液氮中；6个月后复苏细胞，观察细胞的生长形态，台盼蓝染色排除法计数活细胞。 2．在复苏后培养至第8代的细胞中，加入40ng/ml HGF和10ng／mlFGF-4，阴性对照组为不加诱导因子培养的同代细胞，，阳性对照组为HepG2；于加入诱导因子后第7、14、21及28天检测细胞胞浆内CK818、白蛋白及糖原的表达。 3．将类肝细胞与新鲜制备的UCBMSCs共培养；将与类肝细胞同期培养的未诱导细胞和新鲜制备的UCBMSCs共培养作为阴性对照组。于28天后检测UCBMSCs中，CK818和白蛋白的双表达以及糖原的表达。
[Abstract]:Objective:To study the effect of cryopreservation and resuscitation on the growth of human umbilical cord blood mesenchymal stem cells (UCB MSCs), and to study the potential of UCBMSCs after resuscitation to differentiate into hepatocytes induced by the combination of hepatocyte growth factor (HGF) and fibroblast growth factor (FGF-4).And whether hepatoid cells have directional induction to UCBMSCs.Methods:1.The sixth passage UCBMSCs was frozen in liquid nitrogen on the cell culture medium containing 10% dimethyl sulfoxide (DMSO). After 6 months the cells were resuscitated to observe the growth morphology of the cells and the living cells were counted by trypan blue staining exclusion method.2.After resuscitation, 40ng/ml HGF and 10 ng / ml FGF-4 were added to the cells cultured in the 8th passage. The negative control group was the same generation of cells cultured without inducible factor.The expression of CK818, albumin and glycogen in the cytoplasm of the cells was detected on the 21st and 28th day after the induction factor was added to the positive control group.3.Hepatoid cells were co-cultured with freshly prepared UCBMSCs and uninduced cells and freshly prepared UCBMSCs were co-cultured as negative control group.After 28 days, the double expression of CK818 and albumin and the expression of glycogen in UCBMSCs were detected.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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