【摘要】： 本實(shí)驗(yàn)從妊娠早期胎兒肝臟中分離獲得具有肝干細(xì)胞特征的肝臟原始細(xì)胞(hFHSCs),對(duì)其一般生物學(xué)特性及增殖分化機(jī)制進(jìn)行研究。結(jié)果如下:(1)通過聯(lián)合酶消化、重力沉降、單克隆消化及選擇性消化獲得hFHSCs;分離純化方法簡(jiǎn)單易操作,很好地保持了細(xì)胞活力,細(xì)胞純度較高;(2)hFHSCs符合肝干細(xì)胞的基本特征:在體外具有很強(qiáng)的增殖能力;同時(shí)表達(dá)肝系及膽系標(biāo)志物,提示具有雙向分化潛能;核型分析正常且無致瘤性,提示為正常細(xì)胞;(3)hFHSCs可表達(dá)EGF、EGF-R及c-met,缺失表達(dá)HGF,表明HGF通過旁分泌途徑作用與hFHSCs細(xì)胞的受體c-met結(jié)合,從而發(fā)揮其生物學(xué)功能;(4)HGF和EGF對(duì)hFHSCs有明顯的促增殖作用;(5)HGF對(duì)hFHSCs向成熟肝細(xì)胞分化有重要影響,EGF可協(xié)同其發(fā)揮作用;(6)建立了體外誘導(dǎo)分化體系,為解決人源性肝細(xì)胞治療供體短缺問題提供了理論線索;(6)HGF可促進(jìn)hFHSCs細(xì)胞中NF-κB活化,該信號(hào)轉(zhuǎn)導(dǎo)途徑可能是HGF促進(jìn)hFHSCs增殖分化的重要通路。 本研究為獲取人源肝干細(xì)胞提供了一種簡(jiǎn)單易行的方法,同時(shí)在體外建立了有效的向成熟肝細(xì)胞誘導(dǎo)分化的培養(yǎng)體系,而分化機(jī)制的探討為進(jìn)一步優(yōu)化該體系提供了理論基礎(chǔ);整體連續(xù)的研究將為解決供體肝細(xì)胞短缺及最終在臨床實(shí)現(xiàn)肝細(xì)胞和肝干細(xì)胞移植提供實(shí)驗(yàn)依據(jù)和新線索。
[Abstract]:In this study, the biological characteristics and the mechanism of proliferation and differentiation of hepatic stem cell (hFHSCs),) were studied by isolating liver progenitor cells from fetal liver in early pregnancy. The results are as follows: (1) the separation and purification of hFHSCs; by combined enzyme digestion, gravity sedimentation, monoclonal digestion and selective digestion were simple and easy to operate, and the cell viability was well maintained. The cell purity is high; (2) hFHSCs accords with the basic characteristics of hepatic stem cells: it has strong proliferative ability in vitro; it also expresses markers of liver system and bile system, which indicates that it has bidirectional differentiation potential; karyotype analysis is normal and non-tumorigenic. (3) hFHSCs can express EGF,EGF-R and c-met. the deletion of HGF, suggests that HGF binds to hFHSCs receptor c-met through paracrine pathway. (4) HGF and EGF could significantly promote the proliferation of hFHSCs; (5) HGF had an important effect on the differentiation of hFHSCs into mature hepatocytes. (6) the system of inducing differentiation in vitro was established. (6) HGF can promote the activation of NF- 魏 B in hFHSCs cells, and this signal transduction pathway may be an important pathway for HGF to promote the proliferation and differentiation of hFHSCs. This study provides a simple and feasible method for obtaining human hepatic stem cells, and establishes an effective culture system for differentiation into mature hepatocytes in vitro. The study of differentiation mechanism provides a theoretical basis for further optimization of the system. The whole continuous research will provide experimental basis and new clue for solving the shortage of donor hepatocytes and finally realizing hepatocyte and liver stem cell transplantation in clinic.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329

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本文編號(hào)：2253903