MOUSE HEPATOCYTES MIGRATE TO LIVER PARENCHYMA AND FUNCTION INDEFINITELY AFTER INTRASPLENIC TRANSPLANTATION
MOUSE HEPATOCYTES MIGRATE TO LIVER PARENCHYMA AND FUNCTION INDEFINITELY AFTER INTRASPLENIC TRANSPLANTATION
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DOI：
10.1073/pnas.88.4.1217
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发表时间：
1991-02-01
影响因子：
11.1
通讯作者：
WOO, SLC
中科院分区：
文献类型：
--
作者：
PONDER, KP;GUPTA, S;WOO, SLC
肝脏疾病基因治疗的一种方法是从受影响的个体中取出肝细胞，在体外对其进行遗传改变，并将其重新植入受体位点。 虽然将肝细胞返回肝脏本身是有利的，但由于无法区分供体肝细胞和宿主肝细胞，因此从未评估过这种方法的可行性。 为了明确识别移植后的移植肝细胞，并更好地定量它们的数量和肝功能程度，在C57 BL/6背景下产生了两个转基因小鼠系。 第一种方法从相对肝特异性的人α-1-抗胰蛋白酶（hAAT）启动子表达大肠杆菌β-半乳糖苷酶基因，并允许在5-溴-4-氯-3-吲哚基β-D-半乳糖苷染色后容易地鉴定转基因肝细胞;第二个在相同启动子的控制下产生hAAT蛋白，其能够通过测量hAAT的血清水平来定量肝细胞存活和肝功能的维持。 从转基因供体分离的肝细胞通过脾内注射移植到非转基因C57 BL/6受体中。 令人惊讶的是，在移植后2个月，这些细胞的大部分在肝实质内被鉴定，而不是脾。 在移植受体中检测到的高水平血清hAAT稳定> 6个月，表明建立的细胞将无限期存活。 这些结果对肝脏器官发生和肝脏基因治疗具有重要意义。
One approach to gene therapy for hepatic diseases is to remove hepatocytes from an affected individual, genetically alter them in vitro, and reimplant them into a receptive locus. Although returning hepatocytes to the liver itself would be advantageous, the feasibility of this approach has never been evaluated due to the inability to distinguish donor from host hepatocytes. To unambiguously identify transplanted hepatocytes after transplantation, and to better quantitate their number and degree of liver function, two transgenic mouse lines were generated in a C57BL/6 background. The first expresses the Escherichia coli beta-galactosidase gene from the relatively liver-specific human alpha-1-antitrypsin (hAAT) promoter and allows transgenic hepatocytes to be readily identified after 5-bromo-4-chloro-3-indolyl beta-D-galactoside staining; the second produces the hAAT protein under control of the same promoter, which enables hepatocyte survival and maintenance of liver function to be quantitated by measuring the serum levels of hAAT. Hepatocytes isolated from transgenic donors were transplanted into nontransgenic C57BL/6 recipients by intrasplenic injection. Surprisingly, a large fraction of these cells were identified within the liver parenchyma but not the spleen at 2 months after transplantation. The high levels of serum hAAT detected in transplant recipients were stable for > 6 months, suggesting that established cells will survive indefinitely. These results have important implications for liver organogenesis and hepatic gene therapy.