Phenotypic stability of adult rat hepatocytes in primary monolayer culture.
Phenotypic stability of adult rat hepatocytes in primary monolayer culture.
复制标题
DOI：
10.1111/j.1749-6632.1980.tb29518.x
复制
发表时间：
1980-01-01
影响因子：
5.2
通讯作者：
Guzelian, P S
中科院分区：
文献类型：
--
作者：
Bissell, D M;Guzelian, P S
体外成熟肝细胞研究的一个主要进展是开发了用蛋白水解酶分散肝脏和收集完整活细胞的方法。Howard和同事为此引入了细菌胶原酶，“这种方法在完整器官中被胶原酶灌注所取代”，因为这种方法大大增加了活细胞的产量。随后证明，胶原酶分离的细胞，在适当的营养培养基中，可以建立在c~培养。由于灌注技术提供了大量的细胞，制备大规模原代培养成为可能，这种方法引起了人们的注意，因为它涉及到肝实质细胞的均匀制备，并且比其他方法获得的成熟分化肝细胞更能维持细胞活力。原代肝细胞培养物可以用非增殖细胞（来自正常肝脏）制备，并在细胞电镀后4小时内建立（见下文）。这种生理上和时间上与完整肝脏的接近，鼓励了许多关于肝脏特异性功能在培养中的保留的研究——这是过去关于肝脏增殖细胞系的一个经常提出的问题。5.对成年大鼠肝细胞进行初代单层培养的第一次研究关注的是特异性功能的定量，将培养的肝细胞与完整的亲代肝脏进行比较，并在某些情况下证明了特异性功能的降低。几个实验室随后的研究证实，不仅哺乳动物肝细胞培养过程中，两栖动物肝细胞也会发生表型变化。它不同于衰老或生存能力受损的过程；它可能在数量上令人印象深刻，涉及关键的肝脏功能。接下来的讨论将关注各种肝细胞培养系统中表型变化的文献记录，以及有关其预防或逆转的现有信息，以及其对肝细胞培养研究的影响。本综述的范围将主要局限于成熟肝细胞的原代培养，不包括胎儿肝细胞的培养和增殖
A major advance in the study of mature hepatocytes ex vivo was the development of methods for dispersing the liver with proteolytic enzymes and harvesting intact viable cells. Howard and coworkers introduced bacterial collagenase for this purpose,'an approach that was superceded in the intact organ by collagenase perfusion'because of substantially increased yields of viable cells by this method. This was followed by the demonstration that collagenase-isolated he atoc tes, in the appropriate nutrient medium, could be established in c~ lture.~'Because the perfusion technique made available large numbers of cells, preparation of mass primary cultures was possible, an approach that has attracted attention because it involves a homogeneous preparation of hepatic parenchymal cells and also sustains cell viability well beyond that obtained with other methods for mature differentiated hepatocytes.'Primary hepatocyte cultures can be prepared with nonproliferating cells (from normal liver) and are established within 4 h of cell plating (see below). This proximity, both physiologically and temporally, to the intact liver has encouraged numerous studies of the retention of liver-specific function in culture-a problem raised frequently in the past with respect to proliferating cell lines from liver. 5.'The first studies of adult rat hepatocytes in primary monolayer culture were concerned with quantitation of specific function, comparing cultured hepatocytes to the intact parent liver, and demonstrated diminished specific function in some instance^.^'Subsequent studies in several laboratories have established that phenotypic change occurs in primary hepatocyte culture, not only in mammalian but also in amphibian hepatocytes7; it is distinct from a process of senescence or impaired viability; and it may be quantitatively impressive, involving key hepatic functions. The discussion that follows will be concerned with the documentation of phenotypic change in various hepatocyte culture systems, with the available information concerning its prevention or reversal and with its implications for studies using hepatocyte culture. The scope of this review will be limited largely to primary culture of mature hepatocytes, to the exclusion of fetal hepatocyte culture and propagated