Efficient Proliferation and Maturation of Fetal Liver Cells in Three-Dimensional Culture by Stimulation of Oncostatin M, Epidermal Growth Factor, and Dimethyl Sulfoxide
Efficient Proliferation and Maturation of Fetal Liver Cells in Three-Dimensional Culture by Stimulation of Oncostatin M, Epidermal Growth Factor, and Dimethyl Sulfoxide
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DOI：
10.1089/ten.tea.2008.0242
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发表时间：
2009-05-01
影响因子：
4.1
通讯作者：
Miyoshi, Hirotoshi
中科院分区：
文献类型：
--
作者：
Koyama, Toshie;Ehashi, Tomo;Miyoshi, Hirotoshi
为了将胎儿肝细胞（FLCs）作为组织工程生物人工肝脏的细胞来源，使用多孔聚合物支架进行FLCs的三维（3-D）培养，同时进行单层培养作为对照。为使FLCs高效生长分化，先在生长培养基中培养3周，再在分化培养基中培养3周。在这些培养中，将刺激因子（肿瘤抑制素M （OSM）、表皮生长因子（EGF）、肝细胞生长因子（HGF）或二甲基亚砜（DMSO））添加到培养基中，并检查它们的作用。当使用含有OSM和EGF的生长培养基时，EGF与OSM协同促进FLCs的生长。对于FLCs向成熟肝细胞的分化，分化培养基中添加DMSO显著增强了三维和单层培养中白蛋白的分泌，尽管HGF仅在单层培养中有效。显微镜观察证实，FLCs仅在含有DMSO的培养基中呈现肝细胞样形态。综上所述，连续给予含有EGF和OSM的生长培养基和含有DMSO的分化培养基可以有效地诱导3d培养的FLCs生长并向成熟肝细胞分化。
For the purpose of applying fetal liver cells (FLCs) as a cell source to tissue-engineered bioartificial livers, three-dimensional (3-D) cultures of FLCs using a porous polymer scaffold, as well as monolayer cultures as a control, were simultaneously performed. To achieve efficient growth and differentiation, the FLCs were cultured in the growth medium for the first 3 weeks and then cultured in the differentiation medium for 3 more weeks. In these cultures, stimulating factors (oncostatin M (OSM), epidermal growth factor (EGF), hepatocyte growth factor (HGF), or dimethyl sulfoxide (DMSO)) were added to the media, and their effects were examined. When the growth medium containing OSM and EGF was used, EGF stimulated the growth of FLCs synergistically with OSM. For the differentiation of FLCs into mature hepatocytes, DMSO added to the differentiation medium remarkably enhanced albumin secretion in the 3-D and monolayer cultures, although HGF was effective only in the monolayer culture. Microscopic observation proved that FLCs exhibited hepatocyte-like morphology only in the media containing DMSO. In conclusion, successive supply of the growth medium containing EGF and OSM and the differentiation medium containing DMSO efficiently induced the growth of the 3-D cultured FLCs and their differentiation into mature hepatocytes.