MAINTENANCE OF DIFFERENTIATED RAT HEPATOCYTES IN PRIMARY CULTURE
MAINTENANCE OF DIFFERENTIATED RAT HEPATOCYTES IN PRIMARY CULTURE
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DOI：
10.1073/pnas.82.10.3252
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
MUMMAW, J
中科院分区：
文献类型：
--
作者：
ISOM, HC;SECOTT, T;MUMMAW, J
正常的成年大鼠肝细胞保持活力和功能至少43天时，铺在胶原蛋白包被的培养皿和饲料化学成分确定的培养基补充二甲基亚砜（Me 2SO）。通过胶原酶灌注分离并在存在或不存在Me 2SO的情况下培养的肝细胞，通过光学和电子显微镜检查形态学变化;分析白蛋白和其他血浆蛋白的产生，并通过放射自显影法检测DNA合成。Me 2SO处理的细胞在整个培养期间继续产生特异性血浆蛋白;白蛋白产量从接种后第2天至至少第43天一直很高（11-19 μ g/ml培养基/24小时）。超微结构分析表明，Me 2SO处理的肝细胞类似于那些从完整的肝细胞质细胞器和细胞连接的组织。观察Me 2SO形态学和生化效应的最佳浓度为2%（体积/体积）。用Me 2SO补充化学成分确定的培养基能够使分化的肝细胞在培养物中维持较长的时间。
Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were examined by light and electron microscopy for morphological changes; analyzed for the production of albumin and other plasma proteins, and tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 .mu.g/ml of culture medium per 24 h) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). Supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time.