Isolation and long-term culture of human hepatocytes.
Isolation and long-term culture of human hepatocytes.
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发表时间：
1993
期刊：
影响因子：
3.8
通讯作者：
C. Ryan;E. Carter;R. Jenkins;L. Sterling;M. Yarmush;R. Malt;R. Tompkins
中科院分区：
文献类型：
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作者：
C. Ryan;E. Carter;R. Jenkins;L. Sterling;M. Yarmush;R. Malt;R. Tompkins
外科肝脏切除标本的压力控制灌流提高了长期培养研究中人肝细胞的产量。用Dulbecco‘s改良Eagle和Williams E培养液在单层胶原层或双层胶原胶之间培养，评价细胞外基质构型对白蛋白分泌的影响。12例患者的肝细胞培养超过30天，其中5例细胞培养超过2个月。在双凝胶中，细胞呈典型的多边形肝细胞形态，白蛋白分泌量高达65天(p<；0.01)；相反，在单凝胶中，细胞水平铺展，白蛋白分泌在3周内迅速下降。比较不同的培养基配方，白蛋白的最大分泌量出现在5天后，但在杜尔贝科培养液中维持的时间明显更长(p<；0.01)。简单地添加第二层胶原蛋白，形成胶原蛋白三明治，显着稳定和支持人类肝细胞的长期培养。
Pressure-controlled perfusion of specimens of surgical hepatic resection produced improved yields of human hepatocytes for studies of long-term cultures. The effect of an extracellular matrix configuration on albumin secretion was evaluated by culture on a single layer of collagen or between double layers of collagen gel with Dulbecco's modified Eagle and Williams E media. Hepatocytes from 12 patients were maintained for more than 30 days, and in five of 12 experiments cells were cultured beyond 2 months. In the double gels the cells demonstrated typical polygonal liver cell morphology and higher albumin secretion (p < 0.01) up to 65 days; in contrast, in the single gels cells spread horizontally, and albumin secretion declined rapidly within 3 weeks. A comparison of media formulations showed that maximum albumin secretion occurred 5 days later but was maintained significantly longer with Dulbecco's medium (p < 0.01). The simple addition of a second layer of gelled collagen forming a collagen sandwich significantly stabilizes and supports the long-term culture of human hepatocytes.