Profiling the impact of medium formulation on morphology and functionality of primary hepatocytes in vitro.
The characterization of fully-defined in vitro hepatic culture systems requires testing of functional and morphological variables to obtain the optimal trophic support, particularly for cell therapeutics including bioartificial liver systems (BALs). Using serum-free fully-defined culture medium formulations, we measured synthetic, detoxification and metabolic variables of primary porcine hepatocytes (PPHs)--integrated these datasets using a defined scoring system and correlated this hepatocyte biological activity index (HBAI) with morphological parameters. Hepatic-specific functions exceeded those of both primary human hepatocytes (PHHs) and HepaRG cells, whilst retaining biotransformation potential and in vivo-like ultrastructural morphology, suggesting PPHs as a potential surrogate for PHHs in various biotech applications. The HBAI permits assessment of global functional capacity allowing the rational choice of optimal trophic support for a defined operational task (including BALs, hepatocellular transplantation, and cytochrome P450 (CYP450) drug metabolism studies), mitigates risk associated with sub-optimal culture systems, and reduces time and cost of research and therapeutic applications.
Hepatology Laboratory, University of Edinburgh, Chancellor's Building, Royal Infirmary of Edinburgh, EH16 4SB, Scotland, UK.
Leonard J
LJ
Nelson
Philipp
P
Treskes
A Forbes
AF
Howie
Simon W
SW
Walker
Peter C
PC
Hayes
John N
JN
Plevris
D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D015388:Organelles / Q000648:ultrastructure; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000096:biosynthesis; D013552:Swine; D014508:Urea / Q000378:metabolism
D003470:Culture Media; D012709:Serum Albumin; D014508:Urea; D003577:Cytochrome P-450 Enzyme System; D005690:Galactose