Microcarrier culture of hepatocytes in whole plasma for use in liver support bioreactors.
Contact activation of plasma clotting may limit the use of some microcarrier types for hepatocyte attachment in a liver assist device. Activation by seven microcarrier types was studied in plasma containing 2-500 units heparin/mL. Clotting was activated by dextran (Cytodex 1 and 2) and collagen-coated (Cytodex 3) microcarriers at 2-25 units/mL (Cytodex 1) and 2-100 units/mL (Cytodex 2 and 3). There was no activation by polystyrene, gelatin, glass or fibronectin-coated polystyrene microcarriers. Compared with culture medium, incubation of HepG2 cells in plasma did not affect cell viability but increased cell number (56.4 versus 65.1 x 10(4) cells; P less than 0.05) and incorporation of [3H]-amino acids into protein (204913 versus 279624 dpm; P less than 0.05). Polystyrene-attached cells demonstrated time-linear protein synthesis, glucose and 7-ethoxycoumarin metabolism. We conclude that polystyrene-attached hepatocytes maintain viability and metabolic activity in plasma and are of potential use in a liver support bioreactor.
Cunningham
J M
JM
Department of Medicine, Royal Postgraduate Medical School, London, UK.
Hodgson
H J
HJ
D001187:Artificial Organs; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism; D003470:Culture Media; D003584:Cytological Techniques; D005112:Extracorporeal Circulation / Q000295:instrumentation*; D005337:Fibrin / Q000096:biosynthesis; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D010949:Plasma; D013997:Time Factors
D003470:Culture Media; D005337:Fibrin