Engraftment of human hepatocytes in the livers of rats bearing bone marrow reconstructed with immunodeficient mouse bone marrow cells
Engraftment of human hepatocytes in the livers of rats bearing bone marrow reconstructed with immunodeficient mouse bone marrow cells
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DOI：
10.1111/j.1399-3089.2008.00483.x
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发表时间：
2008-07-01
影响因子：
3.9
通讯作者：
Yoshizato, Katsutoshi
中科院分区：
文献类型：
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作者：
Igarashi, Yuka;Tateno, Chise;Yoshizato, Katsutoshi
背景：此前，我们创建了一种嵌合小鼠（人源化小鼠），这是一种严重联合免疫缺陷（SCID）小鼠，其肝脏> 90％重新填充了人（h）肝细胞，可用于测试药物代谢和毒性。以及乙型肝炎病毒和丙型肝炎病毒易感动物模型。然而，它们较小的体型和较小的总血量限制了其用于分析目的，这使得我们开发了一种方法来创建具有h肝细胞再生肝脏的嵌合大鼠。方法：用X射线照射缺乏T细胞的F344裸鼠，并注射来自SCID小鼠（m（SCID））的骨髓细胞（BMC）。通过外周血单核细胞 (PBMC) 的双色流式细胞术分析评估 m(SCID)-BMC 的替代率。 m(SCID)-BMC 重新填充宿主骨髓 (BM) 后，用逆转录酶治疗大鼠。部分肝切除（PHx），并移植从嵌合小鼠中分离出的 5 x 10(6) h 肝细胞。分别采用ELISA、免疫染色和逆转录-PCR检测宿主血液中的h-白蛋白（h-Alb）浓度以及h-肝细胞中肝细胞分化标志物的蛋白和mRNA的表达水平。结果：m(SCID)-BMC成功地在大鼠中进行再增殖，在m-BMC移植后4周，小鼠细胞占宿主（r(nudeF344)）PBMC的百分比达到94%。 h-从嵌合小鼠中分离的肝细胞被移植到m(SCID)-BMC重新填充的大鼠的肝脏中。移植的 h 肝细胞表达 h-Alb 和 h-细胞色素 P450 (CYP) 亚型，并存活并显示正常表型，直到 h-肝细胞移植 (h-HPCT) 后至少 3 周。然而，HPCT 后 4 周，h-Alb 的血液浓度下降，同时出现 r(nudeF344)- 和 m(SCID)- 巨噬细胞，表明由于巨噬细胞的激活而出现 h-肝细胞排斥。结论：我们开发了一种新方法，利用具有由 m(SCID)-BMC 组成的 BM 的大鼠作为肝脏，来创建移植有 h-肝细胞的肝脏的大鼠。主机。这种携带 h 肝细胞的大鼠将成为研究异种移植中涉及的免疫机制和产生具有更高 h 肝细胞再生率的大鼠的有价值的模型。
Background: Previously., we created, a chimeric mouse (humanized mouse), a severe combined immunodeficiency (SCID) mouse whose liver was > 90% repopulated with human (h)-hepatocytes., which are useful for the testing of drug metabolism and toxicity. as well as a hepatitis B virus and hepatitis C Virus-susceptible animal model. However, their small body size and small total blood Volume limited the utilization for analytical purposes, which led us to develop a method to create a chimeric rat bearing h-hepatocyte-repopulated liver.Methods: F344 nude rats devoid of T cells were irradiated with X-rays and injected with bone marrow cells (BMCs) from SCID mice (m(SCID)). The rate of replacement with m(SCID)-BMCs was evaluated by two-color flow cytometry analysis of peripheral blood mononuclear cells (PBMCs). After m(SCID)-BMCs repopulated the host bone marrow (BM), the rats were treated with retrorsine. partially hepatectomized (PHx), and transplanted with 5 x 10(6) h-hepatocytes isolated from the chimeric mice. h-Albumin (h-Alb) concentrations in the host blood and the expression levels of protein and mRNA of hepatocyte differentiation markers in the h-hepatocytes were evaluated by ELISA, immunostaining., and reverse transcription-PCR, respectively.Results: The m(SCID)-BMCs successfully repopulated the rats, the percentage of mouse cells reaching 94% among host (r(nudeF344)) PBMCs at 4 weeks after m-BMC transplantation. h-Hepatocytes isolated from the chimeric mice were transplanted to the liver of the m(SCID)-BMC-repopulated rats. The engrafted h-hepatocytes expressed h-Alb and h-cytochrome P450 (CYP) subtypes and survived showing normal phenotypes until at least 3 weeks post-h-hepatocytes transplantation (h-HPCT). However, the blood concentrations of h-Alb declined at 4 weeks post-HPCT, concomitant with the emergence of both r(nudeF344)- and m(SCID)- macrophages, suggesting the rejection of h-hepatocytes due to the activation of macrophages.Conclusion: We developed a novel method to create a rat that bears the liver engrafted with h-hepatocytes, utilizing a rat with the BM composed of m(SCID)-BMCs as a host. This h-hepatocyte-bearing rat will be a Valuable model for studying the immunologic mechanisms involved in xenogeneic transplantation and for generating rats with higher rates of repopulation with h-hepatocytes.