Deng Xing, Chen Yue-Xiang, Zhang Xin, Zhang Jun-Ping, Yin Chuan, Yue Hai-Yan, Lin Yong, Han Ze-Guang, Xie Wei-Fen
Department of Gastroenterology, Changzheng Hospital, Second Military Medical University, Shanghai, China.
J Cell Physiol. 2008 Oct;217(1):138-44. doi: 10.1002/jcp.21481.
Differentiation of stem cells is tightly regulated by the microenvironment which is mainly composed of nonparenchymal cells. Herein, we investigated effect of hepatic stellate cells (HSCs) in different states on mesenchymal stem cells (MSCs) differentiation. Rat HSCs were isolated and stayed quiescent within 5 days. Primary HSCs were activated by being in vitro cultured for 7 days or cocultured with Kupffer cells for 5 days. MSCs were cocultured with HSCs of different states. Expression of hepatic lineage markers was analyzed by RT-PCR and immunofluorescence. Glycogen deposition was detected by periodic acid-schiff staining. MSCs cocultured with HSC-T6 or Kupffer cell activated HSCs were morphologically transformed into hepatocyte-like cells. Hepatic-specific marker albumin was expressed in 78.3% of the differentiated MSCs 2 weeks after initiation of coculture. In addition, the differentiated MSCs also expressed alpha-fetoprotein, cytokeratin-18, glutamine synthetase and phosphoenolpyruvate carboxykinase. Glycogen deposition was detectable in 55.4% of the differentiated MSCs 6 weeks after initiation of coculture. However, the quiescent HSCs or culture activated HSCs did not exert the ability to modulate the differentiation of MSCs. Moreover, Kupffer cell activated HSCs rather than culture activated HSCs expressed hepatocyte growth factor mRNA. We draw the conclusion that fully activated HSCs could modulate MSCs differentiation into hepatocyte-like cells.
干细胞的分化受到主要由非实质细胞组成的微环境的严格调控。在此，我们研究了不同状态的肝星状细胞（HSC）对间充质干细胞（MSC）分化的影响。分离大鼠HSC，并在5天内保持静止。原代HSC通过体外培养7天或与库普弗细胞共培养5天而被激活。将MSC与不同状态的HSC共培养。通过RT-PCR和免疫荧光分析肝谱系标志物的表达。通过过碘酸希夫染色检测糖原沉积。与HSC-T6或库普弗细胞激活的HSC共培养的MSC在形态上转变为肝细胞样细胞。共培养开始2周后，78.3%的分化MSC表达肝特异性标志物白蛋白。此外，分化的MSC还表达甲胎蛋白、细胞角蛋白-18、谷氨酰胺合成酶和磷酸烯醇丙酮酸羧激酶。共培养开始6周后，55.4%的分化MSC中可检测到糖原沉积。然而，静止的HSC或培养激活的HSC没有发挥调节MSC分化的能力。此外，库普弗细胞激活的HSC而非培养激活的HSC表达肝细胞生长因子mRNA。我们得出结论，完全激活的HSC可以调节MSC分化为肝细胞样细胞。
