TY - JOUR
T1 - Simplified method for cryopreservation of islets using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants
AU - Maruyama, M.
AU - Kenmochi, T.
AU - Sakamoto, K.
AU - Arita, S.
AU - Iwashita, C.
AU - Kashiwabara, H.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2004/5
Y1 - 2004/5
N2 - Cryopreservation is an ideal method for long-term storage of human islets. Dimethyl sulfoxide (DMSO) has been used as an intracellular cryoprotectant. However, because of its toxicity, DMSO has to be added stepwise and diluted stepwise with sucrose. We combined hydroxyethyl starch (HES) as an extracellular cryoprotectant with DMSO to simplify the freeze-thawing procedure. Islets were isolated from the pancreas of beagle dogs by an automated digestion method and Ficoll purification. After overnight culture, the islets were cryogeneically stored using cooling by a programmed freezing system. After 4-week storage in liquid nitrogen, the container was rapidly thawed in a 37°C water bath. The function of the islets was assessed upon static incubation immediately after thawing, showing a recovery rate of 71.16% ± 20.14% and a stimulation index of 1.80 ± 0.78. In conclusion use of HES allowed a decrease in DMSO concentration and simplified the freeze-thawing procedure for islets.
AB - Cryopreservation is an ideal method for long-term storage of human islets. Dimethyl sulfoxide (DMSO) has been used as an intracellular cryoprotectant. However, because of its toxicity, DMSO has to be added stepwise and diluted stepwise with sucrose. We combined hydroxyethyl starch (HES) as an extracellular cryoprotectant with DMSO to simplify the freeze-thawing procedure. Islets were isolated from the pancreas of beagle dogs by an automated digestion method and Ficoll purification. After overnight culture, the islets were cryogeneically stored using cooling by a programmed freezing system. After 4-week storage in liquid nitrogen, the container was rapidly thawed in a 37°C water bath. The function of the islets was assessed upon static incubation immediately after thawing, showing a recovery rate of 71.16% ± 20.14% and a stimulation index of 1.80 ± 0.78. In conclusion use of HES allowed a decrease in DMSO concentration and simplified the freeze-thawing procedure for islets.
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U2 - 10.1016/j.transproceed.2004.04.016
DO - 10.1016/j.transproceed.2004.04.016
M3 - Article
C2 - 15194395
AN - SCOPUS:2942666291
SN - 0041-1345
VL - 36
SP - 1133
EP - 1134
JO - Transplantation Proceedings
JF - Transplantation Proceedings
IS - 4
ER -