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麋鹿耳皮肤成纤维细胞的分离、培养与核型分析

Separation,culture and karyotype analysis of Pere David′s deer (Elaphursu davidianus) ear skin fibroblast cells

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【作者】 赵雪萍张寒莹王子玉许丹孙永成王锋丁玉华

【Author】 ZHAO Xue-ping1,ZHANG Han-ying1,WANG Zi-yu1,XU Dan1,SUN Yong-cheng1,WANG Feng1,DING Yu-hua2(1.Center of Animal Embryo Engineering Technology,Nanjing Agricultural University,Nanjing 210095,China;2.Dafeng Milu National Natural Reserve of Jiangsu,Dafeng 224136,China)

【机构】 南京农业大学动物胚胎工程技术中心江苏大丰麋鹿国家级自然保护区 江苏南京210095江苏南京210095江苏大丰224136

【摘要】 为获得适合于克隆麋鹿的供体细胞,本试验对麋鹿耳皮肤成纤维细胞的分离、培养、纯化、生长特征、核型分析和冷冻保护剂等进行了研究。通过组织块培养法获得麋鹿皮肤成纤维细胞,用胰酶轻度消化后传代3~4次可得到较纯的皮肤成纤维细胞,并经过免疫荧光鉴定;麋鹿染色体数目为2n=68,核型式为2(M)+64(A),XY(A,M);含10%NCS的DMEM培养基最适宜细胞生长;复苏后可获得87.78%的贴壁率。

【Abstract】 In order to get appropriate nuclear donor for Pere David′s deer(Elaphursu davidianus)cloning,its ear skin fibroblast cells were investigated on separation,culture,purification,growth character,cryoprotectant,and karyotype.The Pere David′s deer ear skin fibroblast cells obtained by tissue culture and enzymatic digestion purified fibroblast cells after 3-4 passages,and then they were confirmed by immunofluorescence.The Pere David′s deer choromosome number is 68,and karyotype formula is 2(M)+64(A),XY(A,M).DMEM medium supplemented with 10% NCS was the most appropriate for cell growth.The survival rate was 87.78% after thawing.

【基金】 江苏省自然科学基金项目(BK2003075)
  • 【文献出处】 南京农业大学学报 ,Journal of Nanjing Agricultural University , 编辑部邮箱 ,2008年01期
  • 【分类号】S865
  • 【被引频次】14
  • 【下载频次】336
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