Simultaneous Determination of Coagulative and Fibrinolytic Activities in Rat Hepatocytes
1993 年 5 巻 1 号 p. 39-49
詳細
1993 年 5 巻 1 号 p. 39-49
Coagulative and fibrinolytic activities were simultaneously determined in isolated or cultured rat hepatocytes, since most coagulative and fibrinolytic factors are synthesized by liver parenchymal cells. Rat hepatocytes were obtained by collagenase perfusion. The hepatocytes were suspended in William's E medium containing fetal bovine serum, and cultured in plastic dishes previously coated with collagen for 24, 48 and 72 hr. Isolated and cultured hepatocytes were collected and sonicated, and medium from cultured cells was concentrated by ultrafiltration. Using these hepatocyte suspensions, the protein contents of the fibrin formed by incubation with Ca++ and tranexamic acid (t-AMCHA), which denoted coagulative activity, were stimulated by epinephrine addition. The protein contents of the insoluble residual fibrin after incubation without t-AMCHA, which expressed fibrinolytic activity, were stimulated by urokinase addition. The degrees of coagulative and fibrinolytic activities of the isolated and cultured hepatocytes were similar. In concentrated medium from cultured hepatocytes, coagulative and fibrinolytic activities can be determined by the same method. From these results, a simultaneous determination of coagulative and fibrinolytic activities in hepatocytes and concentrated medium can be utilized asin vitromodel systems to investigate the direct response of hepatocytes.
