TY - JOUR
T1 - Cold- and cryopreservation of dog liver and kidney slices
AU - Fisher, Robyn L.
AU - Hasal, Steven J.
AU - Sanuik, Jeffery T.
AU - Hasal, Katherine S.
AU - Gandolfi, A. J.
AU - Brendel, Klaus
N1 - Funding Information:
We are grateful to Vitron, Inc. for making available tissue slicers, dynamic organ culture incubators, Teflon and titanium rollers, V7 preservation solution as well as coring tools and other assorted slice-related equipment. This study was supported by NIEHS ES 06067 and ES 05790.
PY - 1996/2
Y1 - 1996/2
N2 - The use of tissue slices in culture could decrease the number of animals used in health-related research and decrease experimental variation. This reduction may come about particularly if the methods of cold- and cryopreserving tissue slices are perfected, and one can conduct sequential in vitro experiments into xenobiotic metabolism, organ-specific toxicity, or organ-specific biochemical processes with tissue slices. With this goal in mind, dog liver and kidney slices were placed in cold storage at 0°C using Viaspan (UW), Euro-Collins (EC), Sacks + prostacyclin (SP), and V-7 (V7) cold-preservation solutions for 10 days. Viability was assessed each day by measuring K+ content and protein synthesis after 4 h of incubation in Waymouth + 10% fetal calf serum (FCS). Dog liver slices can be cold-preserved in V7 for up to 7 days using K+ retention as the viability criterion but only up to 4 days using protein synthesis. Dog kidney slices can be cold-preserved in UW, EC, and V7 for up to 10 days using K+ retention, but only V7 could maintain protein synthesis for 10 days. Cryopreserved dog liver and kidney slices retained 63-68% of control viability after 4 h of incubation in FCS. The cryopreservation regimen included using 10% dimethyl sulfoxide in FCS as the cryoprotectant, a freezing rate of 0.5°C/min for liver slices and 12°C/min for kidney slices, and thawing in 37°C FCS. Continued development of cold- and cryopreserving tissue slices could reduce the numbers of animals used and provide accurate and reproducible data.
AB - The use of tissue slices in culture could decrease the number of animals used in health-related research and decrease experimental variation. This reduction may come about particularly if the methods of cold- and cryopreserving tissue slices are perfected, and one can conduct sequential in vitro experiments into xenobiotic metabolism, organ-specific toxicity, or organ-specific biochemical processes with tissue slices. With this goal in mind, dog liver and kidney slices were placed in cold storage at 0°C using Viaspan (UW), Euro-Collins (EC), Sacks + prostacyclin (SP), and V-7 (V7) cold-preservation solutions for 10 days. Viability was assessed each day by measuring K+ content and protein synthesis after 4 h of incubation in Waymouth + 10% fetal calf serum (FCS). Dog liver slices can be cold-preserved in V7 for up to 7 days using K+ retention as the viability criterion but only up to 4 days using protein synthesis. Dog kidney slices can be cold-preserved in UW, EC, and V7 for up to 10 days using K+ retention, but only V7 could maintain protein synthesis for 10 days. Cryopreserved dog liver and kidney slices retained 63-68% of control viability after 4 h of incubation in FCS. The cryopreservation regimen included using 10% dimethyl sulfoxide in FCS as the cryoprotectant, a freezing rate of 0.5°C/min for liver slices and 12°C/min for kidney slices, and thawing in 37°C FCS. Continued development of cold- and cryopreserving tissue slices could reduce the numbers of animals used and provide accurate and reproducible data.
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U2 - 10.1006/cryo.1996.0016
DO - 10.1006/cryo.1996.0016
M3 - Article
C2 - 8812095
AN - SCOPUS:0030074856
SN - 0011-2240
VL - 33
SP - 163
EP - 171
JO - Cryobiology
JF - Cryobiology
IS - 1
ER -