人脐血单个核细胞诱导肝样细胞的研究

发布时间：2018-01-02 05:24

本文关键词：人脐血单个核细胞诱导肝样细胞的研究　出处：《广西医科大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的:分别在体外采用有细胞因子的培养体系和无细胞因子的培养体系诱导培养脐血单个核细胞,观察肝细胞标志物的表达,探讨体外采用细胞因子诱导脐血单个核细胞(MNC)定向分化为肝细胞的可能性。方法:采集正常产妇脐带血,密度梯度离心法分离、纯化脐血单个核细胞(MNC)。分别用肝细胞生长因子(HGF)、成纤维细胞生长因子(FGF)、HGF+FGF、无生长因子4种处理因素进行诱导培养,采用PT-PCR和免疫组化鉴定肝细胞标志物的表达情况。统计学数据用均数±标准差((?)±s)表示,采用SPSS 10.0统计学软件,用单因素方差分析和t检验对资料进行统计分析。结果:1.密度梯度离心法分离的脐带血单个核细胞,活力及纯度＞95%。CD34~+细胞平均为0.96±0.12%。2.有细胞因子培养组诱导后第7和21天分别检测出肝细胞标志物AFP和白蛋白mRNA的表达,第28天检测出CK-18、抗肝细胞抗体的表达。第21天和28天均可检测到糖原染色阳性细胞。无细胞因子培养组始终未检测到肝系细胞标志。3.统计结果显示,HGF+FGF诱导组肝系细胞标志阳性率均高于单独生长因子诱导组(P＜0.05)。单独HGF、单独FGF诱导组间无明显差异(P＞0.05)。结论:1.无细胞因子作用下,人脐带血单个核细胞不能定向分化为肝细胞样细胞。2.HGF、FGF在体外培养体系中均能诱导人脐带血单个核细胞分化为具有肝系细胞表型和功能的细胞。3.HGF、FGF在诱导人脐带血单个核细胞分化为肝样细胞过程中有协同作用。
[Abstract]:Objective: to investigate the expression of hepatocyte markers in umbilical cord blood mononuclear cells induced by cytokine and non-cytokine culture in vitro. Objective: to investigate the possibility of differentiation of cord blood mononuclear cells (MNCs) into hepatocytes by cytokines in vitro. Methods: umbilical cord blood was collected from normal parturient and purified by density gradient centrifugation. The mononuclear cells of cord blood were purified by HGF and FGFs, respectively. The expression of hepatocyte markers was identified by PT-PCR and immunohistochemistry. Using SPSS 10.0 statistical software, the data were analyzed by single factor ANOVA and t-test. Results 1. Cord blood mononuclear cells were isolated by density gradient centrifugation. Activity and purity > 95. CD34 ~. The expression of hepatocyte markers AFP and albumin mRNA were detected on the 7th and 21st day after induction in the cultured group with cytokines. CK-18 was detected on the 28th day. Expression of anti-hepatocyte antibody. Glycogen staining positive cells could be detected on the 21st and 28th days. HGF. The positive rate of hepatocyte markers in FGF induced group was higher than that in single growth factor induced group (P < 0.05), but there was no significant difference between FGF induction group and FGF alone group (P > 0.05). Conclusion 1. Without cytokines, human umbilical cord blood mononuclear cells can not differentiate into hepatocyte-like cells. 2. In vitro, FGF could induce human umbilical cord blood mononuclear cells to differentiate into hepatocytes with phenotype and function. 3. FGF has synergistic effect in inducing human umbilical cord blood mononuclear cells to differentiate into hepatoid cells.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

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