Generation of hybrid hepatocytes by cell fusion from monkey embryoid body cells in the injured mouse liver
Generation of hybrid hepatocytes by cell fusion from monkey embryoid body cells in the injured mouse liver
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DOI：
10.1007/s00418-005-0065-1
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发表时间：
2006-03-01
影响因子：
2.3
通讯作者：
Teraoka, H
中科院分区：
文献类型：
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作者：
Okamura, K;Asahina, K;Teraoka, H
猴胚胎干细胞具有与人类胚胎干细胞相似的特性，可作为临床前研究的替代模型。当培养由猴ES细胞形成的胚状体（EBs）时，观察到许多肝细胞相关基因的表达，包括细胞色素P450（Cyp）3a和Cyp 7a 1。使用抗白蛋白（ALB）、细胞角蛋白-8/18和α 1-抗胰蛋白酶抗体在发育中的EB中对肝细胞进行免疫细胞化学观察。猴胚胎干细胞在体外向肝细胞系分化的潜能促使我们研究猴EB细胞的可移植性。作为评估再增殖潜力的初步方法，我们将EB细胞移植到免疫缺陷型尿激酶型纤溶酶原激活剂转基因小鼠中，这些小鼠经历肝衰竭。移植后，在小鼠肝脏中观察到表达猴ALB的肝细胞集落。荧光原位杂交结果表明，移植的猴EB细胞和受体小鼠肝细胞之间的细胞融合引起的肝细胞再生。相反，未分化的ES细胞移植后，受体肝脏中既没有观察到细胞融合，也没有观察到肝细胞的再增殖。这些结果表明，在发育中的猴EB，但不污染ES细胞的分化细胞，通过与受体小鼠肝细胞融合产生功能性肝细胞，并重新填充损伤的小鼠肝脏。
Monkey embryonic stem (ES) cells have characteristics that are similar to human ES cells, and might be useful as a substitute model for preclinical research. When embryoid bodies (EBs) formed from monkey ES cells were cultured, expression of many hepatocyte-related genes including cytochrome P450 (Cyp) 3a and Cyp7a1 was observed. Hepatocytes were immunocytochemically observed using antibodies against albumin (ALB), cytokeratin-8/18, and alpha 1-antitrypsin in the developing EBs. The in vitro differentiation potential of monkey ES cells into the hepatic lineage prompted us to examine the transplantability of monkey EB cells. As an initial approach to assess the repopulation potential, we transplanted EB cells into immunodeficient urokinase-type plasminogen activator transgenic mice that undergo liver failure. After transplantation, the hepatocyte colonies expressing monkey ALB were observed in the mouse liver. Fluorescence in-situ hybridization revealed that the repopulating hepatocytes arise from cell fusion between transplanted monkey EB cells and recipient mouse hepatocytes. In contrast, neither cell fusion nor repopulation of hepatocytes was observed in the recipient liver after undifferentiated ES cell transplantation. These results indicate that the differentiated cells in developing monkey EBs, but not contaminating ES cells, generate functional hepatocytes by cell fusion with recipient mouse hepatocytes, and repopulate injured mouse liver.