| Project/Area Number |
10470272
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | The university of Tokyo |
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Principal Investigator |
NAKAJIMA Jun University of Tokyo, Department of Cadiothoracic Surgery, Lecturer, 医学部・附属病院, 講師 (90188954)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Makoto University of Tokyo, Department of Cadiothoracic Surgery, Associate, 医学部・附属病院, 助手 (60302684)
ONO Minoru University of Tokyo, Department of Cadiothoracic Surgery, Associate, 医学部・附属病院, 助手 (40270871)
KAWAUCHI Motohiro University of Tokyo, Department of Cadiothoracic Surgery, Lecturer, 医学部・附属病院, 講師 (00152918)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥8,900,000 (Direct Cost: ¥8,900,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥6,800,000 (Direct Cost: ¥6,800,000)
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| Keywords | Cryopreservation of the human tissue / allogenic transplantation / cell viability / rejection / airway epithelial cell / endothelial cell / fibroblast / Interferon gamma / MLR assay / 細胞培養 / interferon-gamma / HLA-class II / 内皮細胞 |
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| Research Abstract |
Background : Cryopreserved human allogenic heart valves and vessels have come to be widely utilized in cardiothoracic surgey. The harvested tissues are usually frozen in a programmed freezer, which enables slow freezing and avoids tissue destruction by the exotic fever around the freezing point. Cryopreserved tracheas have been found to be more easily implanted than fresh tracheas. The cryopreservation procedure seems to decrease alloimmunogenicity. In this study focused the influence of cryopreservation on the allogenicity of cells included in the tracheal tissue, i.e. airway epithelial cells, vascular endothelial cells and, fibroblasts.
Methods : Viability of the cells was determined by colorimetric MTT assay. Allogenic activity was assessed by flow-cytometric assay determining the degree of increase in expression of allogeneic molecules with interferon-gamma (IFN). One-way mixed lymphocyte-stimulator cell culture (MLC) was also made up.
Results : Viability of program-frozen cells was well preserved in each cell line. Expression of HLA-class II on cell surface was evoked by IFN, which ms not deteriorated by cryopreservation except for the airway epithelial cell. The cryopreserved airway cell line was less sensitive to IFN in expressing HLA-class II antigen. MLC of fibroblast and lymphocyte showed no decreased reaction even after the fibroblast was cryopreserved and thawed.
Conclusions : Process of cryopreservation variously modulated the allogeneic immunogenicity of the cells included in the human tracheal tissue. It is controversial whether the cryopreservation is favorable for duration of the allograft. Preservation of the cell viability is essential for durability of the allograft, however, it is also associated with the allogenicity of the graft. Further studies concerning the antigen presenting cells, such as the dendritic cells included in the allograft, would reveal the modulation of allogenicity in cryopreserved tissues.
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