| Project/Area Number |
61480279
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | Fukui Medical School |
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Principal Investigator |
NAKAGAWARA Gizo Fukui Medical School Professor, 医学部, 教授 (10019549)
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| Co-Investigator(Kenkyū-buntansha) |
NOTE Masayuki Fukui Medical School Assistant Professor, 医学部, 助手 (60189412)
KOJIMA Yasuhiko Fukui Medical School Assistant Professor, 医学部, 講師 (00135071)
福島 弥 福井医科大学, 医学部, 助手 (10199218)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1988: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Pancreatic Islet / Cryopreservation / Rapid Cooling Rate / Islet Viability / ラ島再生能 / 移植 / 新生児膵 / 再生能 / 急速凍結保存 / インスリン分泌能 / インスリン含有量 / DNA複製能 / ラ島移植 / 膵ラ氏島 / 膵ラ氏島内分泌能 / 膵ラ氏島移植 |
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| Research Abstract |
Concerning the use of cryopreservation for pancreatic islet storage, there are several factors to consider in order to obtain the best functional presenvation of islet cells. The most obvious one is the cooling rate. In our studies, the isolated islets were cultured free-floating in petri-dish with RPMI 1640 + 10% calf serum in air + 5% CO_2 at 37゜C After 3 days of culture, the islets were cooled in Hanks' solution containing 15% Me_2SO and 10% calf serum at a cooling rate of 25゜C/min to -70゜C using a programmable temperature controller. Finally, these islets were immersed in liquid nitrogen and kept frozen for 2 weeks. The function of cryopreserved islets was compared with that of non-frozen cultured islets (control). Islet viabilities were evaluated by determining glucose-stimulated insulin release, islet contents of insulin and DNA, replicatory capacity of the islet cells in vitro and in vivo, and therapeutic effects of the islets in streptozotocin-induced diabetic recipients. In li
… More
ght microscopic examination of frozen-thawed islets, cryopreserved islets resembled the non-frozen cultured control islets. The basal insuin release from cryopreserved islets were almost the same as that from the control. When challenged with high glucose concentration, the insulin secretory rate of the cryopreserved islets increased more than 3-fold. The DNA content of cryopreserved islets was 0.465 0.067 ng/10 islets, which was about 30% less than the control. Similar observation was made with respect to islet content of insulin. However, the insulin content was expressed on a per DNA besis, there was no difference between cryopreserved and non-frozen control islets. The labelling index of the cryopreserved islets were about 2.04% in vitro and 1.38% in vivo, which were almost the same as those of the controls. Implantation of 1000 cryopreserved syngeneic islets into diabetic recipients led to complete normalization of the hyperglycemia in all of eight hamsters. It is concluded that the fairly rapid cooling rate of 25゜C/min may be useful for cryopreservation of isolated pancreatic islets. Less
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