| Project/Area Number |
03670612
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | Kanazawa Medical University |
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Principal Investigator |
KOJIMA Yasuhiko Kanazawa Medical University, Associate Professor, 医学部, 助教授 (00135071)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAGAWARA Gizo Fukui Medical School, Professor, 医学部, 教授 (10019549)
SAITO Hitoshi Kanazawa Medical University, Assistant Professor, 医学部, 講師 (20178502)
KITA Ichiro Kanazawa Medical University, Assistant Professor, 医学部, 講師 (80195239)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Transplantation / Immunoalteration / Pancreatic islet / Islet cell / Cryopreservation / Pseudoislets |
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| Research Abstract |
For the purpose of pancreatic islet storage, with additional possibility that the populating passenger leukocytes may be reduced, isolated islets were dissociated and then cooled at a cooling rate of 1ﾟC /min to -70ﾟC by using a programmable temperature controller. Then they were stored in liquid nitrogen for 2 weeks. The present paper describes the morphological and biological studies of the islet cells cultured in vitro after freezing-thawing process.
Isolated pancreatic islets from Fischer rats can be dissociated by using EDTA and dispase. These islet cells were also cryopreserved without major loss of their numbers and after thawing they were aggregated into pseudoislets in vitro culture (FCIC). Histological studies of immunostaining with anti-insulin and anti-glucagon antibodies demonstrated the distribution of cells containing these insulin and glucagon hormones, which was the same of that of non-frozen control cultured islets. The test of glucose-stimulating insulin release from the pseudoislets resulted in a response as good as non-frozen control islets.
We concluded that dissociated pancreatic islet cells can be cryopreserved without a loss of their biological abilities. Moreover, there was a significant prolongation of survival time for FCIC in allogenic transplantation, and was different pattern of cell-membrane's protein, included antibody's protein. These results suggested the elimination or alteration of immunogenicity.
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