In vitro differentiation of mouse bone marrow stromal stem cells into hepatocytes induced by conditioned culture medium of hepatocytes
In vitro differentiation of mouse bone marrow stromal stem cells into hepatocytes induced by conditioned culture medium of hepatocytes
复制标题
DOI：
10.1002/jcb.21275
复制
发表时间：
2007-09-01
影响因子：
4
通讯作者：
Xiang, Li-Xin
中科院分区：
文献类型：
--
作者：
Chen, Ye;Dong, Xue-Jun;Xiang, Li-Xin
成体干细胞的分化潜能一直被认为局限于其起源的组织或胚层。然而，最近的研究表明，成体干细胞可能包含比以前想象的更大的潜力。在本研究中，我们研究了小鼠骨髓来源的基质干细胞（BMSSCs）是否能够在体外分化为功能性肝细胞。从小鼠股骨和胫骨中分离BMSSCs，并对这些细胞的中胚层多系分化潜能进行功能表征。为了有效地诱导肝细胞分化，我们设计了一种新的方案，使用肝细胞条件培养基。从小鼠BMSSCs的肝分化进行了检查，在形态学和分子水平的各种测定。形态学上，小鼠BMSSCs诱导后呈圆形、上皮样、双核。在第0、10和20天收获分化的细胞，并通过逆转录-聚合酶链反应（RT-PCR）和免疫细胞化学检查肝细胞特征。我们在mRNA和/或蛋白水平上检测了AFP、HNF-3 β、CK 19、CK 18、ALB、达特和G-6-β，通过在条件培养基中培养的肝细胞样细胞进一步证明了肝细胞的体外功能，包括糖原储存和尿素分泌。此外，分化的细胞移植到肝损伤小鼠部分恢复血清白蛋白水平和显着抑制转氨酶活性。本研究结果提示小鼠BMSSCs在条件培养液中具有向功能性肝细胞样细胞分化的潜能，可作为研究转分化机制的模型系统，也可作为肝脏疾病细胞治疗的细胞来源。J.细胞。102：52-63，2007. (C)2007 Wiley-Liss，Inc.
The differentiation potential of adult stem cells has long been believed to be limited to the tissue or germ layer of their origin. However, recent studies have demonstrated that adult stem cells may encompass a greater potential than once thought. In the present study, we examined whether murine bone marrow derived stromal stem cells (BMSSCs) are able to differentiate into functional hepatocytes in vitro. BMSSCs were isolated from murine femora and tibiae, and the mesodermal multilineage differentiation potentials of these cells were functionally characterized. To effectively induce hepatic differentiation, we designed a novel protocol by using hepatocyte-conditioned medium. Hepatic differentiation from mouse BMSSCs was examined by a variety of assays at morphological and molecular levels. Morphologically, mouse BMSSCs became round and epithelioid, binucleated after induction. Differentiated cells were harvested on Days 0, 10, and 20 and subjected to examination of hepatocyte characteristics by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. We detected AFP, HNF-3 beta, CK19, CK18, ALB, TAT, and G-6-Pase at the mRNA and/ or protein levels, hepatocyte-like cells by culture in conditioned medium further demonstrated in vitro functions characteristic of livercells, including glycogen storage, and urea secretion. Moreover, transplantation of the differentiated cells into liver-injured mice partially restored serum albumin level and significantly suppressed transaminase activity. Our findings indicated the transdifferentiation potential of mouse BMSSCs developing into the functional hepatocyte-like cells by conditioned culture medium and, hence, may serve as a model system for the study of mechanisms involved in the transdifferentiation, and a cell source for cell therapy of hepatic diseases. J. Cell. Biochem. 102: 52-63, 2007. (C) 2007 Wiley-Liss, Inc.