PTH産生ラット上皮小体細胞株の樹立、およびラット同系移植モデルに関する研究

产PTH大鼠甲状旁腺细胞系的建立及大鼠同基因移植模型的研究

• 批准号: 12770852
• 负责人: 徳光 正行
• 金额: $ 1.22万
• 依托单位: Asahikawa Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12770852/
• 关键词: PTH; ラット上皮小体; 初代培養; 継代培養; 細胞株; 同系移植

(1)上皮小体機能亢進症ラット、種大上皮小体の作成前年度同様に、4週齢、雄自然発症性高コレステロール血症ラット(SHCラット)を30週まで自由水摂取下にて飼育。種大上皮小体を得た。(2)ラット上皮小体細胞の分離昨年度同様、摘出した上皮小体の、400Uml collagenase、及び500U/ml dispaseの混合溶液による消化にて、高いviabilityを持つ上皮小体細胞を分離できた。反復施行でも安定した細胞分離結果が得られ、本法が上皮小体細胞分離に適したものであることが、本実験にて確認できた。(3)上皮小体細胞の培養分離された上皮小体細胞は、10%FBS加D-MEM培地にて、各種成長因子の添加、無添加にかかわらず、初代培養においては良好な細胞増殖が見られた。しかし、0.1〜0.125%Trpsine-EDTAを使用した継代では、上皮小体細胞は、各種成長因子の添加、無添加にかかわらず、また培地中のカルシウムやリン濃度の調整を行っても、継代後、上清に分泌されるPTH濃度も低下していくとともに次第に増殖抑制が起こり、最後には上皮小体細胞は死滅、継代不能となっり、本研究当初の目標の細胞株樹立は不可能であった。(4)本研究のまとめ以上のごとく、本研究では、上皮小体細胞の効率のよい分離、初代培養の確立が出来たと考えている。しかしながら、現時点では上皮小体細胞の継代、及び細胞株樹立は不可能であった。今後は、現在、細胞培養の培地に添加する成長因子を変更したり、細胞死(apoptosis)抑制等の観点から、細胞株樹立について検討していこうと考えている。

The main results were as follows: (1) SHC small body hyperactivity syndrome and large epithelial corpuscle were established in the previous year, 4 years old, 4 years old, and male patients with spontaneous hypertrophy, hyperthyroidism, hyperemia, hyperactivity, hyperemia, hyperemia The large epithelial corpuscles are small and small. (2) the epithelial corpuscles were isolated from the same cell last year, and the mixed solution of epithelial corpuscles, 400Uml collagenase, and 500U/ml dispase were isolated. The epithelial corpuscles were isolated from the cells of high viability. The results of antidiazepam cell isolation were satisfactory. In this method, the epithelial corpuscles were isolated and confirmed by this method. (3) the epithelial corpuscles were isolated from the epithelial corpuscles, 10%FBS plus D-MEM culture, various growth factors were added, and the primary culture cells were cultured well. The growth factors were added, added, added and added to the culture medium, and the supernatant and supernatant secreted PTH after culture. After culture, the supernatant was secreted with low concentration of PTH, and the final growth factor was killed. In this study, it was impossible to establish a cell line in the first place. (4) the results of this study are as follows: in this study, the percentage of epithelial corpuscles was isolated, and the primary culture was established in this study. It is not possible to determine the growth of epithelial corpuscles, epithelial corpuscles and cell lines at different times. In the future, cell culture, cell culture, and other growth factors, such as cell death (apoptosis) inhibition, cell growth, cell death, cell growth, cell growth, cell death, cell death, cell death