本文關(guān)鍵詞：人臍血單個(gè)核細(xì)胞誘導(dǎo)肝樣細(xì)胞的研究　出處：《廣西醫(yī)科大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 臍血 單個(gè)核細(xì)胞 分化 肝樣細(xì)胞

【摘要】： 目的:分別在體外采用有細(xì)胞因子的培養(yǎng)體系和無(wú)細(xì)胞因子的培養(yǎng)體系誘導(dǎo)培養(yǎng)臍血單個(gè)核細(xì)胞,觀察肝細(xì)胞標(biāo)志物的表達(dá),探討體外采用細(xì)胞因子誘導(dǎo)臍血單個(gè)核細(xì)胞(MNC)定向分化為肝細(xì)胞的可能性。 方法:采集正常產(chǎn)婦臍帶血,密度梯度離心法分離、純化臍血單個(gè)核細(xì)胞(MNC)。分別用肝細(xì)胞生長(zhǎng)因子(HGF)、成纖維細(xì)胞生長(zhǎng)因子(FGF)、HGF+FGF、無(wú)生長(zhǎng)因子4種處理因素進(jìn)行誘導(dǎo)培養(yǎng),采用PT-PCR和免疫組化鑒定肝細(xì)胞標(biāo)志物的表達(dá)情況。統(tǒng)計(jì)學(xué)數(shù)據(jù)用均數(shù)±標(biāo)準(zhǔn)差((?)±s)表示,采用SPSS 10.0統(tǒng)計(jì)學(xué)軟件,用單因素方差分析和t檢驗(yàn)對(duì)資料進(jìn)行統(tǒng)計(jì)分析。 結(jié)果:1.密度梯度離心法分離的臍帶血單個(gè)核細(xì)胞,活力及純度＞95%。CD34~+細(xì)胞平均為0.96±0.12%。2.有細(xì)胞因子培養(yǎng)組誘導(dǎo)后第7和21天分別檢測(cè)出肝細(xì)胞標(biāo)志物AFP和白蛋白mRNA的表達(dá),第28天檢測(cè)出CK-18、抗肝細(xì)胞抗體的表達(dá)。第21天和28天均可檢測(cè)到糖原染色陽(yáng)性細(xì)胞。無(wú)細(xì)胞因子培養(yǎng)組始終未檢測(cè)到肝系細(xì)胞標(biāo)志。3.統(tǒng)計(jì)結(jié)果顯示,HGF+FGF誘導(dǎo)組肝系細(xì)胞標(biāo)志陽(yáng)性率均高于單獨(dú)生長(zhǎng)因子誘導(dǎo)組(P＜0.05)。單獨(dú)HGF、單獨(dú)FGF誘導(dǎo)組間無(wú)明顯差異(P＞0.05)。 結(jié)論:1.無(wú)細(xì)胞因子作用下,人臍帶血單個(gè)核細(xì)胞不能定向分化為肝細(xì)胞樣細(xì)胞。2.HGF、FGF在體外培養(yǎng)體系中均能誘導(dǎo)人臍帶血單個(gè)核細(xì)胞分化為具有肝系細(xì)胞表型和功能的細(xì)胞。3.HGF、FGF在誘導(dǎo)人臍帶血單個(gè)核細(xì)胞分化為肝樣細(xì)胞過(guò)程中有協(xié)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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 李維,余梅貴,府偉靈;人骨髓基質(zhì)細(xì)胞協(xié)同外源性細(xì)胞因子對(duì)臍血CD_(34)~+細(xì)胞的體外擴(kuò)增作用[J];中國(guó)輸血雜志;2003年02期

2 夏茂林;肝糖原和肌糖原[J];生物學(xué)教學(xué);1997年07期

3 程范軍,鄒萍,楊漢東,余正堂,仲照東;Induced Differentiation of Human Cord Blood Mesenchymal Stem/Pro genitor Cells into Cardiomyocyte-like Cells In Vitro[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)英德文版);2003年02期

4 唐曉鵬,楊旭,張e，

本文編號(hào)：1367872