Comparison of pig hepatocyte isolation using intraoperative perfusion without warm ischemia and isolation of cells from abattoir organs after warm ischemia.
Enzymatic hepatocyte isolation using warm ischemic pig livers from an abattoir was compared with isolation using in situ perfused organs. Using organs from animals of 30 kg body mass (BM), the intraoperative perfusion showed superior results. The use of livers from abattoir pigs of 40-50 kg BM after warm ischemia resulted in a lower yield of hepatocytes and in high rates of injured cells. The mean yield in the intraoperative perfusion group was 68 +/- 11% (wet weight), the maximum yield in the abattoir organ group was 58%. The mean viability in the intraoperative perfusion group was 65 +/- 14% (trypan blue) compared with a maximum viability of 39% in the abattoir liver group. Additional purification by density gradient centrifugation improved the viability of the abattoir liver group to a mean of 95% (trypan blue). The use of pig livers from large abattoir animals required additional purification steps to improve viability since the cell yield is considerably lower than with intraoperative organ perfusion. In general, hepatocyte isolation from abattoir organs is not recommended.
Gerlach
J
J
Chirurgische Klinik, Universitätsklinikum Rudolf Virchow, Freie Universität Berlin, Germany.
Klöppel
K
K
Schön
M R
MR
Brombacher
J
J
Courtney
J M
JM
Unger
J
J
Neuhaus
P
P
D000003:Abattoirs; D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D006358:Hot Temperature; D066298:In Vitro Techniques; D007432:Intraoperative Period; D007511:Ischemia / Q000473:pathology*; D008099:Liver / Q000098:blood supply / Q000166:cytology*; D008297:Male; D010477:Perfusion; D013552:Swine