Rapid generation of functional hepatocyte-like cells from human adipose-derived stem cells.
Rapid generation of functional hepatocyte-like cells from human adipose-derived stem cells.
复制标题
从人类脂肪干细胞中快速生成功能性肝细胞样细胞
DOI：
10.1186/s13287-016-0364-6
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发表时间：
2016-08-05
影响因子：
7.5
通讯作者：
Deng H
中科院分区：
文献类型：
--
作者：
Fu Y;Deng J;Jiang Q;Wang Y;Zhang Y;Yao Y;Cheng F;Chen X;Xu F;Huang M;Yang Y;Zhang S;Yu D;Zhao RC;Wei Y;Deng H
背景肝病是世界范围内导致死亡的主要原因。正交异性肝移植(OLT)是治疗肝功能衰竭的唯一有效方法，但不幸的是，对器官的需求越来越大，其应用受到限制。肝细胞移植是治疗某些肝脏代谢性疾病或急性肝功能衰竭的一种很有前途的替代方法。不幸的是，供体肝脏的缺乏也使得获取足够的可存活的肝细胞用于肝细胞治疗是困难的。目前，这一关键问题仍然缺乏根本解决方案。在这里，我们展示了一种新的非转基因方法，该方法有助于从人脂肪干细胞(HADSCs)中快速生成功能性诱导肝细胞(HHEPS)，为自体肝细胞治疗肝病提供可用的细胞来源。用流式细胞仪检测细胞表面标志物。通过诱导分化为脂肪细胞、骨细胞和软骨细胞来检测其多向分化潜能。结果原代培养的hADSCs呈梭形或多角形，传至第3代后呈成纤维细胞样，95%以上的细胞表达间充质细胞标志物CD29、CD44、CD166、CD105和CD90。HADSC具有向脂肪细胞、骨细胞和软骨细胞多向分化的潜能。我们在体外10天内将hADSCs快速诱导为iHEPs，细胞形态由梭形变为紧密相连的立方形，与肝细胞相似。诱导后，大部分iHEPs共表达白蛋白和α-1抗胰蛋白酶，表达成熟的肝细胞特异性基因，实现了肝细胞的基本功能。结论高纯度hADSCs可在10d内快速诱导分化为有功能的肝细胞样细胞。这些结果为自体肝细胞治疗肝病提供了可用的细胞来源。
BackgroundLiver disease is a major cause of death worldwide. Orthotropic liver transplantation (OLT) represents the only effective treatment for patients with liver failure, but the increasing demand for organs is unfortunately so great that its application is limited. Hepatocyte transplantation is a promising alternative to OLT for the treatment of some liver-based metabolic disorders or acute liver failure. Unfortunately, the lack of donor livers also makes it difficult to obtain enough viable hepatocytes for hepatocyte-based therapies. Currently, a fundamental solution to this key problem is still lacking. Here we show a novel non-transgenic protocol that facilitates the rapid generation of functional induced hepatocytes (iHeps) from human adipose-derived stem cells (hADSCs), providing a source of available cells for autologous hepatocytes to treat liver disease.MethodsWe used collagenase digestion to isolate hADSCs. The surface marker was detected by flow cytometry. The multipotential differentiation potency was detected by induction into adipocytes, osteocytes, and chondrocytes. Passage 3–7 hADSCs were induced into iHeps using an induction culture system composed of small molecule compounds and cell factors.ResultsPrimary cultured hADSCs presented a fusiform or polygon appearance that became fibroblast-like after passage 3. More than 95 % of the cells expressed the mesenchymal cell markers CD29, CD44, CD166, CD105, and CD90. hADSCs possessed multipotential differentiation towards adipocytes, osteocytes, and chondrocytes. We rapidly induced hADSCs into iHeps within 10 days in vitro; the cellular morphology changed from fusiform to close-connected cubiform, which was similar to hepatocytes. After induction, most of the iHeps co-expressed albumin and alpha-1 antitrypsin; they also expressed mature hepatocyte special genes and achieved the basic functions of hepatocyte. Moreover, iHep transplantation could improve the liver function of acute liver-injured NPG mice and prolong life.ConclusionsWe isolated highly purified hADSCs and rapidly induced them into functional hepatocyte-like cells within 10 days. These results provide a source of available cells for autologous hepatocytes to treat liver disease.