A new technique for primary hepatocyte expansion in vitro.
A new technique for primary hepatocyte expansion in vitro.
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DOI：
10.1002/bit.21911
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发表时间：
2008-10-01
影响因子：
3.8
通讯作者：
Yarmush, Martin L.
中科院分区：
文献类型：
--
作者：
Cho, Chenl H.;Berthiaume, Francois;Tilles, Arno W.;Yarmush, Martin L.
目前许多潜在的基于细胞的肝衰竭治疗的应用受到成熟功能肝细胞的低可用性的限制。尽管成人肝细胞在体内具有显著的增殖能力，但在体外增殖成人肝细胞的尝试并不成功。在本研究中，我们研究了共培养细胞类型对肝细胞增殖反应和功能活性的影响。我们首次证明，当与小鼠3T3-J2成纤维细胞共培养时，原代成年大鼠肝细胞具有强大的增殖反应。低密度培养的肝细胞在生长阻滞的3T3-J2成纤维细胞饲养层上的增殖率明显高于在其他四种细胞类型的饲养层上培养的肝细胞。菌落大小的增加与肝细胞功能的增加相关。增殖的肝细胞保留了其形态、表型和功能特征。利用细胞图谱技术，我们发现3T3-J2成纤维细胞通过短距离异型细胞间相互作用刺激肝细胞DNA合成。当在共培养中增殖的肝细胞被收获并进一步在3T3-J2成纤维细胞喂食器或胶原三明治结构中传代培养时，它们的行为与新鲜分离的肝细胞相似。我们得出结论，成年大鼠肝细胞可以体外共培养细胞类型依赖的方式增殖，并且可以通过与3T3-J2成纤维细胞共培养连续繁殖，同时保持其分化特征。我们的研究结果还表明，肝细胞共培养的功能差异的主要原因之一可能是由于肝细胞作为共培养细胞类型的功能的不同增殖反应。本研究为共培养细胞类型和细胞间相互作用在肝脏增殖和功能调节中的作用提供了新的见解。
The current application for many potential cell-based treatments for liver failure is limited by the low availability of mature functional hepatocytes. Although adult hepatocytes have a remarkable ability to proliferate in vivo, attempts to proliferate adult hepatocytes in vitro have been less successful. In this study, we investigated the effect of coculture cell type on the proliferative response and the functional activities of hepatocytes. We show, for the first time, a robust proliferative response of primary adult rat hepatocytes when cocultured with mouse 3T3-J2 fibroblasts. Hepatocytes cultured at low density on growth-arrested 3T3-J2 fibroblast feeder layers underwent significantly higher proliferation rates than when cultured on feeder layers made of four other cell types. Increasing colony size correlated with an increase in hepatocellular functions. The proliferating hepatocytes retained their morphologic, phenotypic, and functional characteristics. Using a cell patterning technique, we found that 3T3-J2 fibroblasts stimulate DNA synthesis in hepatocytes by short-range heterotypic cell–cell interactions. When hepatocytes that proliferated in cocultures were harvested and further subcultured either on 3T3-J2 fibroblast feeders or in the collagen sandwich configuration, their behavior was similar to that of freshly isolated hepatocytes. We conclude that adult rat hepatocytes can proliferate in vitro in a coculture cell type-dependent manner, and can be serially propagated by coculturing with 3T3-J2 fibroblasts while maintaining their differentiated characteristics. Our results also suggest that one of the major reasons for the functional differences in hepatocyte cocultures may be due to the different proliferative responses of hepatocytes as a function of coculture cell type. This study provides new insights in the roles of coculture cell types and cell–cell interactions in the modulation of hepatic proliferation and function.
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影响因子：
15.9
作者：
BISSELL, DM;ARENSON, DM;ROLL, FJ
通讯作者：
ROLL, FJ
影响因子：
13.5
作者：
GOULET, F;NORMAND, C;MORIN, O
通讯作者：
MORIN, O
影响因子：
4.8
作者：
Cho, Cheul H.;Parashurama, Natesh;Yarmush, Martin L.
通讯作者：
Yarmush, Martin L.
影响因子：
2.9
作者：
DUNN, JCY;TOMPKINS, RG;YARMUSH, ML
通讯作者：
YARMUSH, ML
影响因子：
11.4
作者：
FRASLIN, JM;KNEIP, B;GUGUENGUILLOUZO, C
通讯作者：
GUGUENGUILLOUZO, C