Cryopreservation of human pancreatic islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants.
Although widely used, DMSO is toxic for pancreatic islets. We combined hydroxyethyl starch (HES) with DMSO to simplify the procedure of freezing and thawing, and to decrease the toxicity of DMSO. A preclinical study was performed using islets from beagle dogs. After storage for 4 weeks, the islets were thawed and examined. The islet structure was well maintained after thawing. Although the number of the islets decreased to 71.2 +/- 20.1%, the function of the islets was evaluated by static incubation after thawing and showed a 1.80 +/- 0.78 stimulation index. We have introduced this technique for the cryopreservation of human islets from non-heart-beating donors. Twelve cases of human islet cryopreservation were performed. The sample tube of each human cryopreservation was thawed to evaluate the morphology, contamination, and endocrine function. Although fragmentation was observed in five samples (41.6%), the other seven (58.4%) showed a normal structure when evaluated by microscopic and electron microscopic study. The stimulation index (SI) of static incubation deteriorated from 3.37 +/- 3.02 to 1.34 +/- 0.28 after thawing. We divided the thawed islets into two groups: group 1 (n=8), SI > 1.2; group 2 (n=4), SI < 1.2. The group 1 islets showed a higher rate of normal structure (87%) than did group 2 (25%). Moreover, the SI before cryopreservation was 4.01 +/- 3.57 in group 1, which was higher than the SI of 2.11 +/- 0.72 in group 2. Based on the good results from the preclinical study using a large-animal model, this method was introduced for clinical application. Even from the pancreata of non-heart-beating donors, a successful islet cryopreservation was achieved. However, the isolated islets with poor function should not be cryopreserved for transplantation.
Kenmochi
Takashi
T
Department of Surgery, Chiba-East National Hospital, National Hospital Organization, Chiba, Japan. kenmoti@cehprinet.com
Asano
Takehide
T
Maruyama
Michihiro
M
Saigo
Kenichi
K
Akutsu
Naotake
N
Iwashita
Chikara
C
Ohtsuki
Kazunori
K
Suzuki
Akiko
A
Miyazaki
Mariko
M
D000818:Animals; D002102:Cadaver; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D004285:Dogs; D006801:Humans; D006895:Hydroxyethyl Starch Derivatives; D007515:Islets of Langerhans; D016381:Islets of Langerhans Transplantation; D014019:Tissue Donors
D003451:Cryoprotective Agents; D006895:Hydroxyethyl Starch Derivatives; D004121:Dimethyl Sulfoxide