Cryopreservation of rat, dog and human hepatocytes: influence of preculture and cryoprotectants on recovery, cytochrome P450 activities and induction upon thawing.
Several cryopreservation protocols for hepatocytes have been proposed over the past few years, but their effectiveness varies greatly as a function of the characteristics of the method used. One factor in the success of cryopreservation is the quality of cells before freezing. The results suggest that the cryopreservation of hepatocytes in a medium containing polyvinylpyrrolidone (PVP), in addition to DMSO, constitutes a convenient means of long-term storage of hepatocytes for preparing primary cultures to be used in drug metabolism studies. The combined use of the two cryoprotectants is particularly critical for low-viability cell suspensions. An interesting alternative to increase cell viability is the preculture of hepatocytes before cryopreservation. By the use of this procedure, high-quality cells, estimated in terms of post-thaw recovery, viability, adaptation of hepatocytes to culture, drug-metabolizing capability and cytochrome P450 induction, are obtained. Therefore, cryopreserved hepatocytes can provide a regular source of metabolically competent cells for in vitro investigations of the metabolic profile of new drugs and drug-drug interactions in pharmaco-toxicological research.
Gómez-Lechón
M J
MJ
Unidad de Hepatología Experimental, Centro de Investigación, Hospital Universitario La Fe. Avda, Valencia, Spain. gomez_mjo@gva.es
Lahoz
A
A
Jiménez
N
N
Vicente Castell
J
J
Donato
M T
MT
D000818:Animals; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004285:Dogs; D004353:Drug Evaluation, Preclinical; D004789:Enzyme Activation / Q000187:drug effects; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology*; D006358:Hot Temperature; D006801:Humans; D008297:Male; D011205:Povidone / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D003451:Cryoprotective Agents; D003577:Cytochrome P-450 Enzyme System; D011205:Povidone; D004121:Dimethyl Sulfoxide