Growth and functional activities of neonatal and adult rat hepatocytes cultured on fibronectin coated substratum in serum-free medium |
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Authors: | Normand Marceau Micheline Noël Julien Deschênes |
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Institution: | (1) Laboratoire de Biophysique Médicale, le Centre Hospitalier de l’Université Laval, G1V 4G2;(2) le Département de Médecine, Université Laval, Sainte-Foy, Quebec, Canada |
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Abstract: | Summary Hepatocytes isolated from neonatal (NN) and adult (AD) rats were seeded on fibronectin coated substratum and cultured in arginine-free
medium supplemented with various combinations of insulin, dexamethasone, triiodothyronine (T3), albumin, and transferrin, in presence or absence of fibronectin depleted serum (FDS). The main finding is that in response
to certain hormone mixtures, both NN and AD hepatocytes can be stimulated to proliferate, as revealed by an increase in cell
number, a 3H]thymidine incorporation into nuclei, and extractable DNA as well as the appearance of mitotic figures. Moreover, this proliferative
activity is associated with changes in hepatocyte ploidy. However, the proliferative response of NN hepatocytes to hormone
action is much different from that of AD hepatocytes, and the addition of FDS amplifies this activity in NN but inhibits it
in AD hepatocyte cultures. Measurements of tyrosine aminotransferase and lactate dehydrogenase activities indicate a good
preservation of NN and AD hepatocyte functional integrity under certain culture conditions. A good maintenance of albumin
production in NN and AD hepatocyte cultures requires the presence of dexamethasone, whereas theα-fetoprotein production in NN hepatocyte cultures is reduced quite rapidly under most conditions. Noα-fetoprotein is detectable in AD hepatocyte cultures.
Part of this work was presented at the 31st Annual Meeting of the Tissue Culture Association, St. Louis, MO, June 1980. |
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Keywords: | hepatocytes primary cultures fibronectin coating proliferation polyploidy functional activities |
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