A MORPHOLOGICAL-STUDY OF DIFFERENTIATED HEPATOCYTES IN-VITRO
A MORPHOLOGICAL-STUDY OF DIFFERENTIATED HEPATOCYTES IN-VITRO
复制标题
DOI：
10.1016/0270-9139(95)90371-2
复制
发表时间：
1995-07-01
期刊：
影响因子：
13.5
通讯作者：
HEIFETZ, AH
中科院分区：
文献类型：
--
作者：
ARTERBURN, LM;ZURLO, J;HEIFETZ, AH
原代肝细胞培养传统上遇到的一个问题是它们的快速去分化，这不仅反映在肝脏特异性功能的下降，而且反映在去分化的形态上：细胞变平，去极化，并且在体内失去正常肝细胞的许多表面特征。然而，维持原代大鼠肝细胞处于健康和高度分化状态的培养条件最近得到了发展：肝细胞在添加地塞米松和二甲基亚砜（DMSO）的Chee培养基中在胶原包被的Permanox培养皿中培养。除了保留不稳定的肝细胞特异性功能（如P450活性和白蛋白合成）外，这些肝细胞还具有分化的形态。它们的亚细胞细胞器形态正常，在邻近细胞的膜内形成专门的连接和胆管，肌动蛋白纤维聚集在这些胆管表面。这些肝细胞也能合成凝血因子，这些凝血因子聚集成细胞间的纤维蛋白网。总之，这些形态学数据表明，这些肝细胞是极化的，通常具有与肝脏实质细胞非常相似的外观，并且促进肝脏特异性功能保留的相同培养条件对维持生理形态也至关重要。这种分化的形态，包括细胞的极化性质，是在不使用血清或灵活或复杂的细胞外基质的情况下建立的，并显示了细胞结构和组织特异性功能之间的密切联系。
A problem traditionally encountered with primary hepatocyte cultures is their rapid dedifferentiation, which is reflected not only in decreased liver-specific functions, but also in dedifferentiated morphology: the cells flatten, depolarize, and lose many of the surface characteristics of normal hepatocytes in vivo. However, culture conditions that maintain primary rat hepatocytes in a healthy and highly differentiated state were recently developed: the hepatocytes are cultured in Chee's Medium supplemented with dexamethasone and dimethyl sulfoxide (DMSO) on collagen-coated Permanox dishes. In addition to retaining labile hepatocyte specific functions (e.g., P450 activity and albumin synthesis), these hepatocytes also have a differentiated morphology. They have numerous microvilli and are cuboidal and cluster into cords reminiscent of hepatic trabeculae, Their subcellular organelles have normal morphology, and specialized junctions and bile canaliculi form within the membranes of adjacent cells, Actin fibers cluster art these canalicular surfaces. These hepatocytes also synthesize blood clotting factors, which aggregate into fibrin meshworks between cells, Taken together, these morphological data suggest that these hepatocytes are polarized and generally have an appearance very similar to parenchymal cells in the liver, and that the same culture conditions that promote retention of liver-specific functions are also critical to the maintenance of physiological morphology, In contrast to other hepatocyte cultures, this differentiated morphology, including the polarized nature of the cells, is established without the use of serum or flexible or complex extracellular matrices and shows a close link between cellular architecture and tissue-specific function.