ROLE OF PLASMIN INHIBITORS IN GROWTH OF ADULT RAT HEPATOCYTES IN SERUM-FREE PRIMARY CULTURE

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Isolated adult rat hepatocytes attached to flasks in the presence of frbronectin, spread, formed monolayer, and subsequently proliferated in Koga’s medium L under serum-free conditions. When a plasmin inhibitor, aprotinin (Trasylol) was omitted from the medium, hepatocytes tended to shrink after initial spreading, were detached from the substrata and died within 2 or 3 days. Examination of the hepatocyte-conditioned medium revealed the presence of plasmin-like protease activity. Since other plasmin inhibitors also prevented hepatocytes from shrinking and detachment, it is suggested that plasmin-like protease(s) is produced by hepatocytes and this causes detachment of hepatocytes. Thus, it is necessary to supplement a plasmin inhibitor such as aprotinin for successful cultivation of hepatocytes under serum-free conditions.