Classification of the isolated hepatocytes
Nagamori, S.; Fujise, K.; Hasumura, S.; Homma, S.; Sujino, H.; Matsuura, K.; Kameda, H.
Human Cell 1(2): 178-187
1988
ISSN/ISBN: 0914-7470 PMID: 2856499 Document Number: 321526
At this present, enzyme perfusion method is a routine technique to isolate hepatocytes from rat liver for the physiological and pathological experiments. This study described a way of the classification of freshly isolated hepatocytes. First of all, the hepatocytes were fractionated with parenchymal and non-parenchymal cells by low speed centrifugation. And then these cells were subfractionated with a newly developed Percoll linear density gradient method. The fractionated parenchymal cells were divided with cells of periportal and centrilobular areas, respectively. Furthermore, their characteristics were confirmed functionally and morphologically. Non-parenchymal cells (NPC) include Kupffer cells, endothelial cells and fat storing cells (FSC, Ito cells). These isolated NPC are fractionated with a method as mentioned above or centrifugal alutriation method. In this paper, fractionation and classification of Kupffer cells and FSC were discussed with the measurement of fluorescent intensity of vitamin A and the morphological observation of cytoskeleton in culture. Especially, transport of vitamin A into FSC were detected autoradiographically.