Document Detail

Hypothermic preservation of hepatocytes. II. Importance of Ca2 and amino acids.
MedLine Citation:
PMID:  2311409     Owner:  NLM     Status:  MEDLINE    
The importance of the components of a tissue culture media, Leibovitz-15 (L-15), for maintaining viability of hypothermically preserved hepatocytes was analyzed. Hepatocytes isolated from rat livers were incubated at 5 degrees C in an oxygenated environment with continuous shaking (to simulate organ perfusion preservation). L-15 + 5 g% polyethylene glycol (PEG) or variants of this solution were used as the preservation media. After 48 hr of storage, hepatocyte viability was assessed by measuring the release of LDH into the incubation medium and cell volumes were determined. Following 90 min of normothermic incubation (to simulate organ reperfusion), mitochondrial function was measured. Hepatocytes stored in the complete L-15 solution were about 90% viable at the end of 48 hr of storage, while cells stored in a solution containing only the principle electrolytes (PE) lost viability (70% viable). Only the addition of a combination of divalent cations (Ca/Mg) and amino acids was sufficient to maintain viability equivalent to that obtained in the complete L-15 mixture. Hepatocytes suspended in L-15 maintained normal cell volumes (3.85 microliters/mg protein), while cells in the PE solution were swollen with cell volumes of 4.66 microliters/mg protein. Only the addition of Ca/Mg to the PE solution was effective at suppressing cell swelling similar to the complete L-15 media. Both basal and uncoupler-stimulated respiration were depressed in cells stored in the PE solution (15 and 28 nmol O2/min/mg protein) as compared to cells in L-15 (21 and 41 nmol O2/min/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS)
D C Marsh; F O Belzer; J H Southard
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Cryobiology     Volume:  27     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  1990 Feb 
Date Detail:
Created Date:  1990-04-26     Completed Date:  1990-04-26     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1-8     Citation Subset:  IM    
Department of Surgery, University of Wisconsin Hospital, Madison 53792.
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MeSH Terms
Amino Acids
Cell Survival
Cold Temperature
Culture Media
Liver / cytology*,  metabolism
Oxygen Consumption
Rats, Inbred Strains
Tissue Preservation / methods*
Grant Support
Reg. No./Substance:
0/Amino Acids; 0/Culture Media; 7440-70-2/Calcium

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