Cryopreservation of human pancreatic Islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants

Takashi Kenmochi, Takehide Asano, Michihiro Maruyama, Kenichi Saigo, Naotake Akutsu, Chikara Iwashita, Kazunori Ohtsuki, Akiko Suzuki, Manko Miyazaki

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)61-67
Number of pages7
JournalCell Transplantation
Volume17
Issue number1-2
DOIs
Publication statusPublished - 01-01-2008

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Thawing
Dimethyl sulfoxide
Cryopreservation
Dimethyl Sulfoxide
Starch
Islets of Langerhans
Freezing
Poisons
Toxicity
Animals
Contamination
Pancreas
Animal Models
Transplantation
Electrons
Dogs

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

Cite this

Kenmochi, Takashi ; Asano, Takehide ; Maruyama, Michihiro ; Saigo, Kenichi ; Akutsu, Naotake ; Iwashita, Chikara ; Ohtsuki, Kazunori ; Suzuki, Akiko ; Miyazaki, Manko. / Cryopreservation of human pancreatic Islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants. In: Cell Transplantation. 2008 ; Vol. 17, No. 1-2. pp. 61-67.
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abstract = "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.",
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Cryopreservation of human pancreatic Islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants. / Kenmochi, Takashi; Asano, Takehide; Maruyama, Michihiro; Saigo, Kenichi; Akutsu, Naotake; Iwashita, Chikara; Ohtsuki, Kazunori; Suzuki, Akiko; Miyazaki, Manko.

In: Cell Transplantation, Vol. 17, No. 1-2, 01.01.2008, p. 61-67.

Research output: Contribution to journalArticle

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T1 - Cryopreservation of human pancreatic Islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants

AU - Kenmochi, Takashi

AU - Asano, Takehide

AU - Maruyama, Michihiro

AU - Saigo, Kenichi

AU - Akutsu, Naotake

AU - Iwashita, Chikara

AU - Ohtsuki, Kazunori

AU - Suzuki, Akiko

AU - Miyazaki, Manko

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