【摘要】：背景 我國重型肝炎或急性肝功能衰竭發(fā)生率較高,其病理特點是大塊或亞大塊肝細胞壞死,而殘存肝細胞的增殖受到多種因素的抑制,因此依賴病變肝臟自身的能力引發(fā)肝再生困難。 原位肝移植(OLT)是迄今為止治療終末期肝病的有效的方法,但因供肝不足、費用昂貴且需終身使用免疫抑制劑等諸多因素限制了其臨床應用。 肝細胞移植作為輔助治療方法是肝臟移植的重要補充,但肝細胞的來源、數(shù)量、移植后肝細胞在體內(nèi)的增殖和功能等問題尚未完全解決,阻礙著它的發(fā)展,理論上用正常人肝細胞作為細胞移植來源最為理想,但同樣存在供體短缺的問題,且增殖能力較差;異種肝細胞雖然來源廣泛,但又存在免疫排斥、病毒感染特別是逆轉(zhuǎn)錄病毒感染等問題;肝腫瘤細胞株增殖能力強,但存在腫瘤種植風險,且肝細胞功能較差;永生化細胞系應用最多的是SV40 LT基因轉(zhuǎn)染的永生化肝細胞,雖解決了增殖難題,但安全性受到置疑。 人胎肝細胞可分泌細胞生長因子刺激離體細胞生長增殖,組織分化增殖能力強,且免疫原性弱,為較好的細胞來源;臨床資料證實胎兒肝細胞移植在治療肝功能衰竭、先天性酶缺乏等疾病方面有一定的價值,但同樣存在供體缺乏及移植細胞增殖困難等問題。而肝干細胞可以解決這些問題,是因為它具有強大的增殖能力和雙向分化潛能,移植時僅需成熟肝細胞數(shù)量的1/3～1/5,并且胎肝干細胞體積小,更易于從門靜脈注射,移植后易于融入肝板,還有易于大量分離培養(yǎng)和基因操作等優(yōu)點。研究表明處于生長靜止期的成人肝臟中干細胞數(shù)量有限,而胎肝中富含大量肝干細胞,如果選取人胎兒肝干細胞作為細胞移植來源,就可避免未知病原體感染和異種基因?qū)肴梭w,因此,體外成功分離純化人胎兒肝干細胞,并對其進行培養(yǎng)擴增無疑具有非常重要的意義。 本實驗對人胎兒肝干細胞體外分離培養(yǎng)擴增做了初步探索,旨在尋找對其進
[Abstract]:Background the incidence of severe hepatitis or acute liver failure in China is high. The pathological characteristics of severe hepatitis or acute liver failure are large or submassive hepatocytes, while the proliferation of residual hepatocytes is inhibited by many factors. Therefore, it is difficult to regenerate the liver by relying on the ability of the diseased liver itself. Orthopaedic liver transplantation (OLT) is an effective method for the treatment of end-stage liver disease so far, but its clinical application is limited by many factors, such as insufficient donor liver, high cost and lifelong use of immunosuppressive agents. As an auxiliary therapy, liver cell transplantation is an important supplement to liver transplantation, but the source and quantity of hepatocytes, the proliferation and function of hepatocytes in vivo have not been completely solved, which hinders its development. In theory, normal human hepatocytes are the most ideal source of cell transplantation, but there is also a shortage of donors, and the proliferation ability is poor. Although heterogeneic hepatocytes come from a wide range of sources, there are some problems, such as immune rejection, viral infection, especially retrovirus infection, and the proliferation ability of liver tumor cell lines is strong, but there is a risk of tumor implantation, and the function of hepatocytes is poor. Immortalized cell lines are most widely used in immortalized hepatocytes transfected with SV40 LT gene. Although the proliferation problem has been solved, the safety of immortalized cell lines has been questioned. Human fetal hepatocytes can secrete cell growth factor to stimulate the growth and proliferation of isolated cells, the ability of tissue differentiation and proliferation is strong, and the immunogenicity is weak, so it is a good cell source. Clinical data confirmed that fetal liver cell transplantation has certain value in the treatment of liver failure, congenital enzyme deficiency and other diseases, but there are also some problems, such as lack of donor and difficulty in proliferation of transplantation cells. Liver stem cells can solve these problems because they have strong proliferation and bidirectional differentiation potential, only 1 鈮，

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