Simplified method for cryopreservation of islets using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants.
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 degrees 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.
Maruyama
M
M
Department of Surgery, Sakura National Hospital, Chiba, Japan. maruyama@snh.hosp.go.jp
Kenmochi
T
T
Sakamoto
K
K
Arita
S
S
Iwashita
C
C
Kashiwabara
H
H
D000818:Animals; D001331:Automation; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide; D004285:Dogs; D006895:Hydroxyethyl Starch Derivatives; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects
D003451:Cryoprotective Agents; D006895:Hydroxyethyl Starch Derivatives; D004121:Dimethyl Sulfoxide