Hepatocyte culture utilizing porous polyvinyl formal resin maintains long-term stable albumin secretion activity.
To investigate the effects of culture conditions on the maintenance of metabolic functions of cultured hepatocytes, long-term hepatocyte culture lasting 20 days was performed under two different culture conditions, i.e. stationary cultures utilizing porous polymer (polyvinyl formal (PVF) resin) as a substratum and conventional monolayer dish cultures without PVF. Metabolic activities specific to hepatocytes were evaluated in terms of ammonia metabolism, urea synthesis, and albumin secretion. Concerning ammonia metabolic and urea synthetic activities, no significant differences in maintenance of these activities were found between the two culture conditions, and these activities rapidly decreased with the elapse of the culture period, especially during the early stage of the experiments. However, after day 10, these activities in the stationary cultures were maintained at a slightly more favorable level than in the monolayer cultures. On the other hand, compared with ammonia metabolism and urea synthesis, stable and well-maintained albumin secretion of hepatocytes (60% of the activity in day 1) was exhibited in the stationary culture experiments, despite that this particular activity under the monolayer culture condition gradually reduced to a very low level (5.7% of that on day 1) at the end of the culture. From the morphological observations, hepatocytes immobilized in the PVF resin revealed individual spherical shapes without forming multicellular aggregation, and it was suggested that this characteristic structure contributed to good albumin secretion of hepatocytes. In conclusion, the advantages of the hepatocyte culture technique utilizing PVF resin over the conventional dish culture in maintaining some representative metabolic function specific to hepatocytes were clarified.
Miyoshi
H
H
Department of Biomedical Engineering, University of Tsukuba, Ibaraki, Japan.
Ookawa
K
K
Ohshima
N
N
D000418:Albumins / Q000378:metabolism*; D000641:Ammonia / Q000378:metabolism; D002478:Cells, Cultured; D003188:Composite Resins / Q000737:chemistry*; D003470:Culture Media / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry*; D006490:Hemostatics / Q000737:chemistry*; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D011142:Polyvinyl Alcohol / Q000737:chemistry*; D016062:Porosity; D014508:Urea / Q000378:metabolism
D000418:Albumins; D003188:Composite Resins; D003470:Culture Media; D006490:Hemostatics; D005557:Formaldehyde; C027490:polyvinyl alcohol formaldehyde foam; D000641:Ammonia; D014508:Urea; D011142:Polyvinyl Alcohol