Hepatocyte differentiation of mesenchymal stem cells from human adipose tissue in vitro promotes hepatic integration in vivo
Hepatocyte differentiation of mesenchymal stem cells from human adipose tissue in vitro promotes hepatic integration in vivo
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DOI：
10.1136/gut.2008.154880
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发表时间：
2009-04-01
期刊：
影响因子：
24.5
通讯作者：
Christ, B.
中科院分区：
文献类型：
--
作者：
Aurich, H.;Sgodda, M.;Christ, B.
目的：研究人脂肪组织源性间充质干细胞（hAT-MSCs）在体外分化或未分化为肝细胞样细胞时的肝整合情况。方法和结果：从腹膜或皮下脂肪组织中分离的细胞表达间充质干细胞表面标记物，并具有多谱系分化特征。在有利于肝细胞分化的条件下，hAT-MSCs在体外获得了肝细胞功能，包括尿素形成、糖原合成、细胞色素P450酶活性，以及肝细胞特异性氨甲酰磷酸合成酶、白蛋白和细胞色素P450型3A4 （CYP3A4）转录物的表达。绿色荧光蛋白的转基因表达出现在肝细胞分化过程中，由胞质磷酸烯醇丙酮酸羧激酶基因的肝细胞特异性启动子驱动，但由泛素基因启动子组成。将人AT-MSCs移植到免疫缺陷的Pfp/Rag2（-/-）小鼠的肝脏中，事先进行肝细胞分化或不进行肝细胞分化。供体来源的人细胞主要在肝小叶的门静脉周围区域植入小鼠宿主肝脏。它们表达HepPar1和白蛋白，这是分化的人肝细胞的典型特征，在其他阴性的小鼠肝背景下。与未分化的细胞相比，体外预分化为肝细胞样细胞的hAT-MSCs移植效率更高。结论：体外将人间充质干细胞从脂肪组织预分化为肝细胞样细胞有助于体内长期功能性肝整合。
Objective: The hepatic integration of human adipose tissue derived mesenchymal stem cells (hAT-MSCs) in vivo with or without prior differentiation to hepatocyte-like cells in vitro was investigated.Methods and results: Cells, isolated either from peritoneal or subcutaneous adipose tissue, expressed mesenchymal stem cell surface markers and featured multiple lineage differentiation. Under conditions favouring hepatocyte differentiation, hAT-MSCs gained hepatocytic functions in vitro including urea formation, glycogen synthesis, cytochrome P450 enzyme activity, and expression of hepatocyte-specific transcripts of carbamoylphosphate synthetase, albumin and cytochrome P450 type 3A4 (CYP3A4). Transgenic expression of green fluorescent protein emerged upon hepatocyte differentiation when driven by the hepatocyte-specific promoter of the cytosolic phosphoenolpyruvate carboxykinase gene but was constitutive from the ubiquitin gene promoter. Human AT-MSCs were transplanted into livers of immunodeficient Pfp/Rag2(-/-) mice with or without prior hepatocyte differentiation in vitro. Donor-derived human cells engrafted in the mouse host liver predominantly in the periportal region of the liver lobule. They expressed HepPar1 and albumin, typical features of differentiated human hepatocytes, in the otherwise negative mouse liver background. Engraftment was significantly more efficient using hAT-MSCs pre-differentiated to hepatocyte-like cells in vitro as compared with undifferentiated cells.Conclusions: Pre-differentiation of human MSCs from adipose tissue into hepatocyte-like cells in vitro facilitates long term functional hepatic integration in vivo.