Effect of patient, operative and isolation factors on subsequent yield and viability of human hepatocytes for research use.
It is widely accepted that the model of choice for pharmacotoxicological studies are human hepatocytes. There is therefore a demand for these cells, but quality must be maintained for their widespread use. We present a retrospective review of the isolation of hepatocytes from both surgically resected tissue and livers rejected for transplantation, and evaluated patient, operative and isolation variables to ascertain which may affect the viability and yield of cells. Seven clinically rejected whole livers and 60 surgically resected specimens (from two distinct operating centres) were isolated. For surgically resected tissue we found that decreasing age, securing the perfusing cannulae with suture rather than reforming Glissons capsule with glue and steatotic livers improved viability. No significant correlation could be found with pre-operative blood results, disease, type of operation, presence or absence of Pringle manoeuvre, weight of tissue isolated, time of digestion with collagenase and cold ischaemic time. There was a reduction in mean yield and viability when hepatocyte isolations were performed in livers rejected for transplant, compared to surgically resected tissue although this did not reach significance. Human hepatocytes can be successfully and consistently isolated from surgically resected tissue and appear to be superior to those isolated from rejected for transplant livers. From our study, there are few parameters that significantly affect the quality of isolated hepatocytes, which increases the possible pool of tissue that hepatocytes can be isolated from.
Lloyd
T D R
TD
Department of Surgery, Leicester General Hospital, Gwendolen Road, Leicester LE5 4PW, UK. tdlloyd@ntlworld.com
Orr
S
S
Patel
R
R
Crees
G
G
Chavda
S
S
Vadyar
H
H
Berry
D P
DP
Sherlock
D
D
Dennison
A R
AR
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002452:Cell Count; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D005260:Female; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012189:Retrospective Studies; D014019:Tissue Donors