Comparison of Porcine Hepatocytes with Human Hepatoma (C3A) Cells for Use in a Bioartificial Liver Support System

Abstract

Cells from primary porcine hepatocytes (PPH) and the immortalized human hepatoma cell line C3A are both used in bioartificial liver support systems (BALSS). In this work the viability and metabolic capacity of PPH and C3A cells cultured in different media were compared. Also, because the cells come into direct or indirect contact with human blood components in BALSS, the effects of human complement on survival and functions of the cells was evaluated. For short-term culture, maintenance of PPH viability was essential for retention of P450IA1 activity (r = 0.882, p < 0.01) and effective ammonia clearance (r = -0.791, p < 0.01). When cell viability was below 60% P450IA1 activity could not be recorded and nitrogen elimination activity significantly diminished. In contrast to PPH, ammonia levels were markedly increased for C3A cells in all culture media tested (p < 0.01). Ammonia increase correlated with C3A viability (r = 0.896, p < 0.05). PPH metabolic function was superior to that of the C3A cell line when evaluated by P450IA1 activity, ammonia removal, and amino acid metabolism. When PPH were incubated in human plasma (HP) or human serum (HS) there was rapid and irreversible deterioration of viability occurring within 9 h. This toxic effect could be prevented by the inactivation of complement. When sodium citrate dissolved in dextrose was added to medium, there was considerable damage to both PPH and the C3A cell line. However, there was no demonstrable toxic effect when hepatic cells of either type were exposed to heparin. We conclude that PPH cultivated in complement-inactivated HP or HS are to be preferred to C3A for clinical application of BALSS, and that heparin should be preferred for anticoagulation in BALSS.

Which hepatocyte will it be? Hepatocyte choice for bioartificial liver support systems

“…However, this still remains a theoretical concern because no evidence exists of transmission in trials performed to date. 17 The 2 basic types of human hepatocyte lines are those derived from human tumors 18,19 and those created by immortalizing human hepatocytes. [20][21][22] Tumor-derived line.…”

“…26 However, compared with primary porcine hepatocytes, the C3A cell line has lower levels of P-450 IA1 activity, ammonia removal, and amino-acid metabolism. 19 Increasing evidence shows that continuous human liver cell lines derived from liver cancers may not conserve all aspects of drug metabolism found in normal cells, further limiting their efficacy. 27 Immortalized cell lines.…”

Cells for bioartificial liver devices: The human hepatoma‐derived cell line C3A produces urea but does not detoxify ammonia

“…Two such cell lines, HepG2 and the HepG2 subclone C3A, are commonly used (Kelly and Darlington, 1989). Previous studies have shown that C3A cells, in particular, are capable of producing urea, a finding which has been generally interpreted as indicating the presence of a fully functional urea cycle capable of ammonia detoxification via this specific pathway, and has led to their use in clinically trialled bioartificial devices (Chen et al, 2003;David et al, 2004;Ellis et al, 1996;Filippi et al, 2004;Wang et al, 1998).…”

Current development of bioreactors for extracorporeal bioartificial liver (Review)

“…However, compared to primary porcine hepatocytes, C3A has a lower metabolism ability, particularly to ammonia. 31 Mavri-Damelin et al 32 used the labeled N 15 H 4 CL to detect the ammonia removal capacity of C3A and found that the urea generated did not contain the labeled N 15 . By further analysis of reverse transcription polymerase chain reaction ͑RT-PCT͒ and western blotting, the enzymes, ornithine carbamoyl-transferase, and arginase for urea cycle were lacking in C3A, which shows that C3A cells cannot perform the normal ammonia metabolism, instead they produce ammonia through the pathway outside the urea cycle.…”