VIABLE PORCINE HEPATOCYTES FROM SLAUGHTERHOUSE ORGANS

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.