人胎儿肝脏干细胞体外分离培养及增殖分化机制研究
发布时间：2018-10-05 15:26
【摘要】： 本实验从妊娠早期胎儿肝脏中分离获得具有肝干细胞特征的肝脏原始细胞(hFHSCs),对其一般生物学特性及增殖分化机制进行研究。结果如下:(1)通过联合酶消化、重力沉降、单克隆消化及选择性消化获得hFHSCs;分离纯化方法简单易操作,很好地保持了细胞活力,细胞纯度较高;(2)hFHSCs符合肝干细胞的基本特征:在体外具有很强的增殖能力;同时表达肝系及胆系标志物,提示具有双向分化潜能;核型分析正常且无致瘤性,提示为正常细胞;(3)hFHSCs可表达EGF、EGF-R及c-met,缺失表达HGF,表明HGF通过旁分泌途径作用与hFHSCs细胞的受体c-met结合,从而发挥其生物学功能;(4)HGF和EGF对hFHSCs有明显的促增殖作用;(5)HGF对hFHSCs向成熟肝细胞分化有重要影响,EGF可协同其发挥作用;(6)建立了体外诱导分化体系,为解决人源性肝细胞治疗供体短缺问题提供了理论线索;(6)HGF可促进hFHSCs细胞中NF-κB活化,该信号转导途径可能是HGF促进hFHSCs增殖分化的重要通路。 本研究为获取人源肝干细胞提供了一种简单易行的方法,同时在体外建立了有效的向成熟肝细胞诱导分化的培养体系,而分化机制的探讨为进一步优化该体系提供了理论基础;整体连续的研究将为解决供体肝细胞短缺及最终在临床实现肝细胞和肝干细胞移植提供实验依据和新线索。
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R329
本文编号：2253903
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R329
【参考文献】
相关期刊论文 前2条
1 肖家诚,朱延波,朱上林,李勤裕,RuckP,AdamA,KaiserlingE;肝细胞肝癌中卵圆细胞的组织学与超微结构研究[J];临床与实验病理学杂志;2000年03期
2 龚加庆,方驰华,李雅,田伏洲;卵圆细胞参与实验性肝癌形成过程的研究[J];中华外科杂志;2004年05期
，本文编号：2253903
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2253903.html
最近更新
教材专著
