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目的:探讨干细胞样肝原始细胞集落体外分化的潜能。方法:分离孕13.5 d的C57BL/6小鼠胎肝组织细胞并培养,第2天出现圆形或椭圆形干细胞样肝原始细胞集落,随机挑取边界清楚的细胞集落分别培养,利用倒置相差显微镜连续观察细胞生长过程中的形态学变化并用显微摄影记录结果,最后利用免疫细胞化学分析鉴定分化后细胞的性质。结果:干细胞样肝原始细胞集落在体外培养时可形成类似肝小叶的形状,集落中央大部分细胞分化成双核细胞,集落周边细胞仍为单核细胞,但体积明显增大,整个集落中的细胞呈放射状排布。分化后,细胞群中的部分细胞表达成熟肝细胞标志白蛋白(albumin),少部分细胞表达胆管上皮细胞标志细胞角蛋白19(cytokeratin19,CK19),但所有细胞均不表达低分化肝细胞标志α-甲胎蛋白(α-fetoprotein,AFP)。结论:干细胞样肝原始细胞在体外具有多向分化能力,可分化为表达成熟肝细胞和胆管上皮细胞标志的细胞。
Objective: To investigate the potential of stem cell-like hepatoblastoid cells to differentiate in vitro. METHODS: C57BL / 6 mice fetal liver cells were isolated and cultured at 13.5 days of gestation. On the second day, round or oval stem cell-like hepatocyte colonies appeared. Colonies with clear border were randomly selected and cultured. The cells were cultured with inverted phase contrast microscope The morphological changes during the growth of the cells were observed continuously and the results were recorded by photomicrography. Finally, the characteristics of the differentiated cells were identified by immunocytochemical analysis. Results: The stem cell-like hepatoblast colony formed hepatic lobule-like shape when cultured in vitro. Most of the cells in the central part of the colony differentiated into dinuclear cells. The surrounding cells in the colony were still mononuclear cells, but the volume was significantly increased. The cells in the whole colony Radial arrangement. After differentiation, some of the cells in the cell population expressed mature hepatocyte albumin (albumin), while a few expressed cytokeratin 19 (CK19), but none of the cells expressed low grade hepatocyte marker α - Alpha-fetoprotein (AFP). CONCLUSIONS: Stem-like hepatoblasts have a multidirectional differentiation capacity in vitro and can differentiate into cells expressing mature hepatocytes and biliary epithelial cells.