タイトル | ウシ胎仔脳細胞の初代培養と凍結保存技術の開発 |
---|---|
担当機関 | (現 |
研究期間 | 1995~1997 |
研究担当者 | |
発行年度 | 2000 |
要約 | ウシ胎仔脳を材料として,神経細胞,アストログリア,ミクログリア等を初代培養する技術を開発した。また,胎仔脳組織を液体窒素中で凍結保存した後でも,ほぼ同様に初代培養が可能であった。 |
要約(英語) | Procedures for primary cultures and cryopreservation of bovine brain cells were established as in vitro experimental systems, to study the responses of bovine brain cells to neuropathogenic agents. Brain cells were dissociated by papain from the cerebellum of bovine fetuses at 90-120 days of age, and were cultured in different media. In medium containing 1% fetal bovine serum (FBS), neuronal cells were maintained and they formed clusters on glial and fibroblastic cell sheets. In medium containing 10% FBS, the proportion of neurons decreased, and fibroblastic and microglial cells dominated. In serum-free medium containing epidermal growth factor, the highest neuronal proportion was obtained. Optimal cryoperservation conditions for the brain tissues were investigated by changing the concentrations of DMSO and FBS. Brain cells could be cultured from cryopreserved tissue with only slightly reduced growth profiles and varying cell proportions in comparison to those prepared from fresh tissue. (Department of Molecular Biology and Immunology, National Institute of Agrobiological Sciences (NIAS) Tel:+81-298-38-7801) References: 1) A. Hashimoto, T. Onodera, H. Ikeda and H. Kitani: Isolation and characterisation of fetal bovine brain cells in primary culture. Res. Vet. Sci. 69: 39-46 (2000) 2) I. Yamane, H. Kitani, T. Kokuho, T. Shibahara , M. Haritani, T. Hamaoka, S. Shimizu, M. Koiwai, K. Shimura and Y. Yokomizo: The inhibitory effect of interferon gamma and tumor necrosis factor alpha on intracellular multiplication of Neospora caninum in primary bovine brain cells. J. Vet. Med. Sci. 62: 347-351 (2000) 3) H. Kitani, M. Yamakawa and H. Ikeda: Preferential infection of neuronal and astroglia cells by Akabane virus in primary cultures of fetal bovine brain. Vet. Microbiol. 73: 269-279 (2000) |
背景・ねらい | ウシ胎仔に大脳欠損等の異常を引き起こすウイルスが知られているが,これらのウイルスの影響を詳細に解析するための中枢神経系細胞の初代培養技術は,ウシではまだ確立されていない。そこで,ウシ胎仔脳を材料として,神経細胞,アストログリア,ミクログリア等を初代培養する技術の開発を目指した。また,この初代培養技術をより広範な研究に適用可能とするために,ウシ胎仔脳組織の凍結保存技術についても検討した。 |
成果の内容・特徴 |
|
成果の活用面・留意点 |
|
図表1 | ![]() |
図表2 | ![]() |
カテゴリ |