Formation of the Hepatocyte Spheroid and Roles of Plasminogen Activator
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概要
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For culturing hepatocytes, we generally use a matrix, collagen on which the cells proliferate and become confluent monolayer. Recently, a modified proteoglycan, chondroitin sulfatephosphatidylethanolamine (CS-PE), in stead of collagen, has been developed, and made possible a novel culture forming a multicellular assembling, spheroid. The spheroid is found to be highly differentiated, since it produces plasminogen and fibrinogen as well as albumin much more than the monolayer does, and it has an ability of being induced tyrosine aminotransferase by hormones. The mechanism with which hepatocytes form spheroid is unknown. Especially, it is not yet known how plasminogen activator (PA) plays in the spheroid or in the process of its formation. Our present study on the fibrinolytic factors revealed that the spheroid was in fairly hypofibrinolytic; t-PA activity in the conditioned media was one-tenth of that of monolayer (11U/106 cells versus 116U/106 cells), and the specific activity of t-PA was around one-fifth of monolayer. In addition, during the culture the mRNA for t-PA decreased in the spheroid, while it increased in monolayer. On the other hand, the mRNA for the PA inhibitor type 1 (PAI-1) was expressed higher in the spheroid. To form spheroid, addition of an anti-t-PA antibody to the media were not hampering although other protease inhibitors, including trypsin and plasmin inhibitors were of quite hampering and the cells remained monolayer. Thus, in the spheroid formation of hepatocytes, the cells would be in fibrinolytically inactive, however, they would require some proteolytic enzyme including plasmin.
- 一般社団法人 日本血栓止血学会の論文
一般社団法人 日本血栓止血学会 | 論文
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