Repopulation of athymic mouse liver by cryopreserved early human fetal hepatoblasts
Repopulation of athymic mouse liver by cryopreserved early human fetal hepatoblasts
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DOI：
10.1089/hum.2004.15.1219
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发表时间：
2004-12-01
影响因子：
4.2
通讯作者：
Weber, A
中科院分区：
文献类型：
--
作者：
Mahieu-Caputo, D;Allain, JE;Weber, A
肝细胞移植是治疗严重肝病的一种有前途的替代肝移植的方法。然而，这种方法受到供体器官短缺和成人肝细胞固有局限性的阻碍。为了研究成人肝细胞所面临的大多数障碍是否可以通过使用人胎儿成肝细胞来克服，我们开发了一种方法来分离、培养和冷冻保存发育早期（妊娠11-13周）的人肝脏成肝细胞。在体外表征细胞的特异性标志物的表达，并在体内表征其移植到无胸腺小鼠后的增殖和分化潜力。大多数细胞（80-90%）具有双能表型，表达细胞角蛋白8/18、白蛋白和CK 19。它们在培养物中自发增殖，并被表达β-半乳糖苷酶的逆转录病毒有效转导（90%）。移植后，冷冻保存的细胞移植到无胸腺小鼠的肝脏中并增殖，导致高达10%的再增殖。移植的细胞表达分化的成体肝细胞的标志物，包括白蛋白、α（1）-抗胰蛋白酶、细胞色素P450 3A 4和α-谷胱甘肽-S-转移酶。当在移植前进行逆转录病毒转导时，它们在体内表达转基因。总之，早期人胎肝母细胞在无胸腺小鼠肝脏中移植、增殖和成熟，而无需调节供体。
Transplantation of hepatocytes is a promising alternative to liver transplantation for the treatment of severe liver diseases. However, this approach is hampered by the shortage of donor organs and intrinsic limitations of adult hepatocytes. To investigate whether most of the hurdles faced with adult hepatocytes could be surmounted by the use of human fetal hepatoblasts, we have developed a method to isolate, transduce, and cryopreserve hepatoblasts from human livers at an early stage of development (11-13 weeks of gestation). Cells were characterized in vitro for expression of specific markers, and in vivo for their proliferation and differentiation potential after transplantation into athymic mice. Most of the cells (80-90%) harbored a bipotent phenotype, expressing cytokeratins 8/18, albumin, and CK19. They proliferated spontaneously in culture and were efficiently transduced by a beta-galactosidase-expressing retrovirus (90%). After transplantation, cryopreserved cells engrafted into the liver of athymic mice and proliferated, resulting in up to 10% repopulation. Engrafted cells expressed markers of differentiated adult hepatocytes including albumin, alpha(1)-antitrypsin, cytochrome P450 3A4, and alpha-glutathione-S-transferase. When retrovirally transduced before transplantation they expressed the transgene in vivo. In summary, early human fetal hepatoblasts engraft, proliferate, and mature in athymic mouse liver, without conditioning the donor.