| 野牦牛成纤维细胞的分离培养与传代 |
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| 引用本文： | 郭志林,杨永梅,赵明德,于军,景佳莹,庞礴,郭松长,赵新全. 野牦牛成纤维细胞的分离培养与传代[J]. 中国畜牧兽医, 2013, 40(12): 156-160 |
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| 作者姓名： | 郭志林 杨永梅 赵明德 于军 景佳莹 庞礴 郭松长 赵新全 |
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| 作者单位： | 1. 青海民族大学化学与生命科学学院, 青海西宁 810007;2. 中国科学院西北高原生物研究所, 青海西宁 810008;3. 青海民族大学公共管理学院, 青海西宁 810007 |
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| 基金项目： | 教育部科学技术研究重点项目（211192）；青海民族大学高层次人才科研项目（2012G002）；财政部战略生物资源科技支撑专项（KSCXZ-YW-Z-0951）。 |
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| 摘 要： | 为了提高野牦牛成纤维细胞分离培养效果，研究了组织保存方法、原代培养方法、培养液类型、添加细胞生长因子和冷冻保存液配方对野牦牛体细胞分离培养与传代的作用。结果表明，PBS、D/F12和TCM199适合野牦牛皮肤组织的保存，D/F12和TCM199培养液适合组织块培养，组织块成活率达到（86.29±4.62）%，组织块法适合野牦牛成纤维细胞的原代培养；D/F12培养液培养野牦牛成纤维细胞效果较好，群体倍增时间和平台期密度分别为38.47 h和2.075×106/mL，正常二倍体核型百分率为84.33%；表皮生长因子（EGF）和碱性成纤维细胞因子（bFGF）产生了较明显的促增殖作用，平台期细胞密度明显增加；2种冷冻保存液（D/F12添加胎牛血清和二甲亚砜及胎牛血清添加二甲亚砜）均能用于野牦牛细胞冷冻，细胞存活率分别是87.33%和89.00%。
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| 关 键 词： | 野牦牛 细胞培养 成纤维细胞 细胞生长因子 |
| 收稿时间： | 2013-06-09 |
Isolation,Culture and Subculture of Skin Fibroblast Cells from Bos mutus |
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| GUO Zhi-lin,YANG Yong-mei,ZHAO Ming-de,YU Jun,JING Jia-ying,PANG Bo,GUO Song-chang,ZHAO Xin-quan. Isolation,Culture and Subculture of Skin Fibroblast Cells from Bos mutus[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(12): 156-160 |
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| Authors: | GUO Zhi-lin YANG Yong-mei ZHAO Ming-de YU Jun JING Jia-ying PANG Bo GUO Song-chang ZHAO Xin-quan |
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| Affiliation: | 1. Academy of Chemistry and Life Science, Qinghai University for Nationality, Xining 810007, China;2. Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, China;3. Academy of Public Management, Qinghai University for Nationality, Xining 810007, China |
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| Abstract: | For improving the efficiency of isolation and culture of fibroblast cells from Bos mutus, effects on the isolation culture and subculture of Bos mutus somatic cells by organize preservation method, primary culture method, culture media type,and cryopreservation media formula were assessed. The results showed that PBS, D/F12 and TCM199 media adapted to preserve the skin organize of Bos mutus. D/F12 and TCM199 media could be used to the culture of Bos mutus organize pieces, and the survival rate of organize pieces reached (86.29±4.62)%. So organize pieces culture could be successfully used to primary culture of Bos mutus fibroblast cells. Better effect was achieved by the culture of Bos mutus fibroblast cells in D/F12 media, and population doubling time as well as plateau phase density of the cells were 38.47 h and 2.075×106/mL, respectively. The percentage of normal diploid karyotype of F7 cells was 84.33% in the D/F12 culture media. Obvious proliferation effect on the cells was produced with the supplement of EGF and bFGF into culture media, and plateaued phase density of the cells obviously rised. Two cryopreservation media (D/F12 supplemented with fetal bovine serum and DMSO, fetal bovine serum supplemented with DMSO) could be used to cryopreservation of Bos mutus cells, and the survival rates of the cells were 87.33% and 89.00%, respectively. |
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| Keywords: | Bos mutus cell culture fibroblast cell cell growth factor |
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