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Long‐Term continuous culture of hepatocytes in a packed‐bed reactor utilizing porous resin
Author(s) -
Miyoshi Hirotoshi,
Yanagi Kennichi,
Ohshima Norio,
Fukuda Hideki
Publication year - 1994
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260430713
Subject(s) - perfusion , ammonium , urea , packed bed , monolayer , chemistry , chromatography , metabolism , biochemistry , organic chemistry , medicine
Abstract As part of our attempt to develop a hybrid artificial liver support system using cultured hepatocytes, we investigated the long‐term metabolic function of hepatocytes incubated in a packed‐bed type reactor using reticulated polyvinyl formal (PVF) resin as a supporting material. Long‐term (up to 1 week) perfusion culture experiments using the packed‐bed reactor (20 mm i.d.) loaded with 500 PVF resin cubes (mean pore size 250 μm, 2 × 2 × 2 mm), together with conventional monolayer culture experiments as controls, were performed in serum‐free or serum‐containing medium. Ammonium metabolism and urea synthesis activities were evaluated quantitatively based on reaction kinetic analyses. Initial rates of ammonium metabolism and urea‐N synthesis, as well as GPT enzyme activities, were adopted as indexes of the metabolic performance of the reactor and activities of the cultured hepatocytes. When serum‐free medium was used in the perfusion cultures, ammonium metabolic and urea‐N synthetic rates showed significant decay with elapse of the culture period, being less than 10% of those measured on day 1. This loss of activity was more prominent in the perfusion culture than in the monolayer cultures using this medium. In contrast, when serum‐containing medium was used, approximately 50% of these activities obtained on day 1 were maintained even at the end of the cultures both in the perfusion and monolayer culture experiments. We concluded that the packed‐bed reactor using PVF resin enabled high‐density culture of hepatocytes, and showed a satisfactory ability to maintain the metabolic function of immobilized hepatocytes for relatively long periods of up to 1 week. This type of reactor is thus considered to represent a breakthrough in overcoming the difficulties involved in the development of a hybridtype artificial liver support system. © 1994 John Wiley & Sons, Inc.