2013 Fiscal Year Final Research Report
Development of simple storage methods for donor cells such as freeze-drying for bovine somatic cell nuclear tramsfer
| Project/Area Number |
23580396
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | Kinki University |
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Principal Investigator |
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| Project Period (FY) |
2011 – 2013
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| Keywords | ウシ / 体細胞クローン / ドナー細胞 / 簡易保存 / 簡易凍結 / 凍結乾燥 / トレハロース |
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| Research Abstract |
Simple storage methods for donor cells used for somatic cell nuclear transfer were examined. First, we examined whether live cells can be retrieved from simple frozen tissues. Bovine several tissues were frozen at -20 oC, -80 oC and -196 oC. After thawing tissues, live cells were obtained from ear tissues frozen at -80 oC even after 6 months. Next, nuclear status of somatic cells after freeze-drying was examined. Trehalose was introduced into cells taken from ear tissues by digitonin, then the cells were freeze-dried. Fragmentation of nuclear DNA was examined by a Comet assay method. DNA fragmentation of freeze-dried cells treated with 0 or 20g/ml digitonin and 0.4M trehalose was similar to that of untreated live cells.
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