caa a22 4500 477080960 CHVBK 20180405111449.0 cr unu---uuuuu 170330e19960101xx s 000 0 eng 10.1007/BF02722991 doi (NATIONALLICENCE)springer-10.1007/BF02722991 The restoration of the functions of serially passaged calf hepatocytes by spheroid formation [Elektronische Daten] [Fumiko Karikusa, Yoshio Sawasaki] Summary: A serial cultivation system of hepatocytes was established for the first time using calf liver as a cell source and, repeating passage of more than 30 cumulative population doublings (PDs), was obtained in the presence of long-acting ascorbic acid derivative (L-ascorbic acid 2-phosphate) and epidermal growth factor. The complete purification of hepatocytes was achieved by repeating ethylenediaminetetraacetic acid (EDTA) treatment, by which hepatocytes were easily detached from the culture dish, leaving most of the nonparenchymal cells on the dish. As the population cumulatively doubled, the cell density and albumin-synthesizing ability decreased gradually, and doubling time has exceeded 120 h at about 30 cumulative PDs. In serially passaged cells, the hepatocyte-specific histochemical and biochemical markers—including glucose-6-phosphatase, ornithine carbamoyltransferase, glutamate hydrogenase, and ammonia-metabolizing activities—have been lost after 20 cumulative PDs. However, when these passaged cells were allowed to form spheroids, the morphologic and biochemical characteristics of hepatocytes have rapidly been restored to levels comparable to those in younger generations. Because no extrinsic factor was needed for this restoration, three-dimensional cell-cell interaction would be indispensable for the differentiation of the hepatocytes. The routine serial cultivation of hepatocytes and their redifferentiation by spheroid formation will be useful for studying metabolism, gene regulation, and transplantation of hepatocytes. Society for In Vitro Biology, 1996 hepatocytes nationallicence serial cultivation nationallicence spheroid nationallicence redifferentiation nationallicence cell-cell interaction nationallicence Karikusa Fumiko Department of Biochemistry, National Defense Medical College, 3-2, Namiki, Tokorozawa, 359, Saitama, Japan aut Sawasaki Yoshio Department of Biochemistry, National Defense Medical College, 3-2, Namiki, Tokorozawa, 359, Saitama, Japan aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/1(1996-01-01), 30-37 1071-2690 32:1<30 1996 32 11626 https://doi.org/10.1007/BF02722991 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02722991 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Karikusa Fumiko Department of Biochemistry, National Defense Medical College, 3-2, Namiki, Tokorozawa, 359, Saitama, Japan aut NATIONALLICENCE 700 1- Sawasaki Yoshio Department of Biochemistry, National Defense Medical College, 3-2, Namiki, Tokorozawa, 359, Saitama, Japan aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/1(1996-01-01), 30-37 1071-2690 32:1<30 1996 32 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer