Comparison of University of Wisconsin and ET-Kyoto preservation solutions for the cryopreservation of primary human hepatocytes.
Primary human hepatocytes are clinically used for transplantation or in bioartificial liver devices for the treatment of patients with liver failure. We aimed to assess whether an organ preservation solution containing trehalose, namely ET-Kyoto solution (ETK), could improve human liver cell viability when used for cryopreservation in comparison to the University of Wisconsin solution (UW). Our study showed beneficial effects of ETK when used in combination with other cryoprotectants on the viability of thawed hepatocytes. Indeed, no significant difference was seen between the viability of freshly isolated cells and cryopreserved cells when cryopreserved with ETK combined with other cryoprotectants. In contrast, a significant decrease of viability was observed in cells cryopreserved with UW or ETK combined with dimethysulfoxide (DMSO) only, or with UW combined with other cryoprotectants, compared to freshly isolated cells. No significant difference was observed between the four different groups of cryopreserved hepatocytes with regards to cell recovery rate or cell attachment after thawing. However, a significant decrease in cell metabolic activity was found in cells cryopreserved with UW 10% DMSO compared to the other groups. In conclusion, our study confirms the beneficial effect of ETK for the cryopreservation of human hepatocytes in combination with other cryoprotective agents.
Illouz
S
S
Department of Hepatobiliary and Pancreatic Surgery, Leicester General Hospital, Leicester, UK. severine.illouz@uhl-tr.nhs.uk
Nakamura
T
T
Webb
M
M
Thava
B
B
Bikchandani
J
J
Robertson
G
G
Lloyd
D
D
Berry
D
D
Wada
H
H
Dennison
A
A
D000241:Adenosine / Q000494:pharmacology; D000328:Adult; D000368:Aged; D000493:Allopurinol / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D005942:Gluconates / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D006498:Hepatectomy; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000473:pathology; D006801:Humans; D006895:Hydroxyethyl Starch Derivatives / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008113:Liver Neoplasms / Q000473:pathology / Q000601:surgery; D008297:Male; D008875:Middle Aged; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D010710:Phosphates / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology; D014021:Tissue Preservation / Q000379:methods; D014199:Trehalose / Q000494:pharmacology
C087059:ET-Kyoto solution; D005942:Gluconates; D006895:Hydroxyethyl Starch Derivatives; D007328:Insulin; D019188:Organ Preservation Solutions; D010710:Phosphates; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D014199:Trehalose; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose