Purification and characterization of a growth factor from rat platelets for mature parenchymal hepatocytes in primary cultures.
A growth factor (HGF) stimulating DNA synthesis of adult rat hepatocytes in primary culture was found in rat platelets. HGF was purified from rat platelets to homogeneity by a three-step procedure: stimulation of its release from platelets by thrombin, cation-exchanger fast protein liquid chromatography on a Mono S column, and heparin-Sepharose chromatography. HGF was clearly distinguishable from the platelet-derived growth factor (PDGF) by fast protein liquid chromatography. HGF was a heat- and acid-labile cationic protein that was inactivated by reduction with dithiothreitol. Its molecular mass was estimated to be 27 kDa by NaDodSO4/PAGE and its amino acid composition was very different from that of PDGF. The purified HGF stimulated DNA synthesis in adult rat hepatocytes at 2 ng/ml and was maximally effective at 20 ng/ml; its effect was additive or synergistic with those of insulin and EGF, depending on their combinations. HGF did not stimulate DNA synthesis of Swiss 3T3 cells, while PDGF did not stimulate that of hepatocytes. Thus, HGF showed clearly different cell specificity from PDGF in its growth-promoting activities. These findings indicate that HGF is a growth factor in platelets for mature hepatocytes.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001792:Blood Platelets / Q000032:analysis / Q000502:physiology*; D001798:Blood Proteins; D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006133:Growth Substances / Q000097:blood* / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D015850:Interleukin-6; D008099:Liver / Q000502:physiology*; D008115:Liver Regeneration; D008970:Molecular Weight; D011506:Proteins / Q000494:pharmacology; D051381:Rats
D000596:Amino Acids; D001798:Blood Proteins; D006133:Growth Substances; D007328:Insulin; D015850:Interleukin-6; D011506:Proteins; D004815:Epidermal Growth Factor