1995 Fiscal Year Final Research Report Summary
Experimentary Studies on Cryopreservation of Dissociated Pancreatic Islet Cells in Relation to Maintenance of Biological Function
| Project/Area Number |
05671055
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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 | Fukui Medical School |
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Principal Investigator |
NAKAGAWARA Gizo Fukui Medical School. The First Department of Surgery. Professor, 医学部, 教授 (10019549)
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| Co-Investigator(Kenkyū-buntansha) |
HIROSE Kazuo Fukui Medical School. School Hospital Faculty of Medicine. The First Department, 医学部附属病院, 講師 (00181199)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Cryopreservation / Islet cell / Pseudoislet / Immunoalteration / Transplantation |
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| Research Abstract |
Recent studies have shown that a prolongation of allograft survival can be obtained by a pretreatment of pancreatic islets to reduce passenger leukocytes prior to transplantation. For the purpose of islet storage, with the additional possibility that the populating passenger leukocytes may be reduced, isolated pancreatic 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 liqid 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 Fisher rats can be dissociated completely by using EDTA and dispase. These islet cells were also cryopreserved without major loss of their numbers and after thawing they were aggregated into pseudo-islets 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 the controls (CI : non-frozen cultured islet). When the insulin content was expressed on a per DNA basis there were no differences between the FCIC and CI,suggesting that the insulin content per islet cell was not altered after these process.
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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