A modified enzymatic isolation technique for the successful harvesting
of porcine liver cells from slaughterhouse organs is introduced. Dige
stion of the left medial liver lobe (n=74) resulted in 1.0+/-0.3 x 10E
7 Viable hepatocytes per gram tissue and an overall yield of 1.92+/-0.
5 x 10E9 cells per isolation (viability: 93+/-2%). Morphological integ
rity of hepatocytes in long term immobilization culture systems was as
sessed by light and electron microscopic follow up. Stable DNA-content
s and low alanine-amino-transferase (ALAT) release were measured after
early culture adaptation. Urea production under NH4Cl, Albumin secret
ion, total bile acid synthesis and a follow up of 7-ethoxicoumarin o-d
eethylase (ECOD) activity demonstrated functional activity and mainten
ance of Type IA1 cytochrome P450 (CYP450) dependent metabolism in cult
ured hepatocytes for at least 10 days. In contrast to published result
s, where hepatocytes have been mainly isolated from anaesthetized anim
als we could not see any disadvantages in the use of liver cells from
slaughterhouse organs. Hepatocytes from slaughtered pigs represent an
unlimited, reliable and inexpensive resource of viable cell material f
or all fields of applied research, including investigations on hepatic
metabolism or the development of hybrid artificial liver support devi
ces. The technique could also help to reduce animal experiments for me
re cell harvesting purposes. We conclude that - wherever possible - po
rcine hepatocytes should be obtained from slaughterhouse organs instea
d of ex vivo cell isolation.