Extended expression of differentiated function in primary cultures of adult liver parenchymal cells maintained on nitrocellulose filters. I. Induction of phosphoenolpyruvate carboxykinase and tyrosine aminotransferase.
Extended expression of differentiated function in primary cultures of adult liver parenchymal cells maintained on nitrocellulose filters. I. Induction of phosphoenolpyruvate carboxykinase and tyrosine aminotransferase.
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在硝化纤维滤膜上维持的成人肝实质细胞的原代培养物中分化功能的扩展表达。
DOI：
10.1016/0014-4827(78)90488-3
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发表时间：
1978
影响因子：
3.7
通讯作者：
R. Bonney
中科院分区：
文献类型：
--
作者：
C. Savage;R. Bonney
建立了一种新的支持系统，可以在单层培养中长期维持未分裂的成年大鼠肝实质细胞。肝细胞附着在Millipore （MP）过滤器上，作为表达分化肝细胞功能的自由漂浮培养物维持长达13天。在MP过滤器上培养8天后，肝细胞仍能诱导3- 4倍的酪氨酸转氨酶和10- 15倍的磷酸烯醇丙酮酸羧激酶。与传统的培养皿（如玻璃或塑料培养皿）相比，使用浮动MP过滤器支持肝细胞的优点是：(1)培养肝细胞的功能寿命延长一倍，允许4-8天完成实验；(2)能够快速、方便地将细胞从一组培养条件转移到另一组培养条件；(3)切片可以从一个过滤器中切割，允许从单个培养中获得多个样品；(4)含有细胞的滤光片可以在不失去细胞表面方向的情况下进行处理，这是采用放射自显影和/或电子显微镜等技术时的重要考虑因素；(5)这种培养技术可以很容易地适用于共培养实验，以便直接检测一种细胞类型的分泌产物对另一种细胞类型的生物学和生化效应。
A new support system has been developed which provides long-term maintenance of non-dividing adult rat liver parenchymal cells in monolayer cultures. The hepatocytes, attached to Millipore (MP) filters, are maintained as free-floating cultures which express differentiated liver cell functions for up to 13 days. After 8 days of culture on MP filters, the hepatocytes are still capable of inducing tyrosine aminotransferase 3- to 4-fold and phosphoenolpyruvate carboxykinase 10-to 15-fold. The advantage of using floating MP filters to support the hepatocytes over the more conventional culture supports such as glass or plastic dishes are: (1) the functional lifespan of cultured hepatocytes is doubled, permitting experiments requiring 4–8 days to complete; (2) it permits rapid and easy transfer of cells from one set of culture conditions to another; (3) sections can be cut from one filter permitting multiple samples from a single culture; (4) the filters containing the cells can be processed without losing the orientation of cell surfaces, an important consideration when employing techniques such as autoradiography and/or electron microscopy; and (5) this culture technique can readily be adapted for co-cultivation experiments in order to directly examine biological and biochemical effects of secreted products of one cell type on another.