Yamazaki Shintaro, Miki Kenji, Hasegawa Kiyoshi, Sata Masataka, Takayama Tadatoshi, Makuuchi Masatoshi
Third Department of Surgery, Nihon University School of Medicine, Tokyo, Japan.
J Hepatol. 2003 Jul;39(1):17-23. doi: 10.1016/s0168-8278(03)00150-8.
BACKGROUND/AIMS: The plasticity of bone marrow cells (BMCs) is shown by their ability to differentiate into mesenchymal as well as endodermal and ectodermal lineages. Transdifferentiation of BMCs into hepatocytes has also been demonstrated, both in vitro and in vivo. In the present study we investigated the effects of liver nonparenchymal cells (NPCs) and sera from liver failure patients (HSLF) on the in vitro transdifferentiation of murine BMCs into hepatocytes.
Liver NPCs from wild-type mice, and 5-azacytidine-treated BMCs from green fluorescence protein transgenic mice, were cocultured in medium containing HSLF in combination with several cytokines. Hepatocyte-specific gene expression in BMCs was identified by immunocytochemistry and reverse transcription-polymerase chain reaction.
Bone marrow cell-derived hepatocyte-like colonies appeared after several days of coculture in medium containing HSLF, oncostatin M (OSM) and hepatocyte growth factor (HGF). These colonies expressed hepatocyte-specific genes. Transdifferentiation was enhanced by 5-azacytidine treatment, and by HSLF, OSM and HGF. It did not take place when the BMCs were separated from the NPCs in a dual chamber dish, or cultured with other mesenchymal cells.
Direct interaction of murine BMCs with liver NPCs, as well as soluble factors in the HSLF and a demethylating agent, strongly stimulate transdifferentiation into hepatocytes.
背景/目的:骨髓细胞(BMCs)的可塑性表现为它们能够分化为间充质细胞以及内胚层和外胚层谱系细胞。BMCs向肝细胞的转分化在体外和体内均已得到证实。在本研究中,我们调查了肝非实质细胞(NPCs)和肝功能衰竭患者血清(HSLF)对小鼠BMCs体外转分化为肝细胞的影响。
将野生型小鼠的肝NPCs与经5-氮杂胞苷处理的绿色荧光蛋白转基因小鼠的BMCs,在含有HSLF并联合几种细胞因子的培养基中共同培养。通过免疫细胞化学和逆转录-聚合酶链反应鉴定BMCs中肝细胞特异性基因的表达。
在含有HSLF、制瘤素M(OSM)和肝细胞生长因子(HGF)的培养基中共同培养几天后,出现了骨髓细胞来源的肝细胞样集落。这些集落表达肝细胞特异性基因。5-氮杂胞苷处理以及HSLF、OSM和HGF可增强转分化。当BMCs与NPCs在双室培养皿中分离或与其他间充质细胞一起培养时,转分化未发生。
小鼠BMCs与肝NPCs的直接相互作用,以及HSLF中的可溶性因子和一种去甲基化剂,强烈刺激其向肝细胞的转分化。