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

作　　者：赵雪萍[1] 张寒莹[1] 王子玉[1] 许丹[1] 孙永成[1] 王锋[1] 丁玉华 

机构地区：[1]南京农业大学动物胚胎工程技术中心,江苏南京210095 [2]江苏大丰麇鹿国家级自然保护区,江苏大丰224136

出　　处：《南京农业大学学报》2008年第1期67-71,共5页Journal of Nanjing Agricultural University

基　　金：江苏省自然科学基金项目(BK2003075)

摘　　要：为获得适合于克隆麇鹿的供体细胞，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；麇鹿染色体数目为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。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 nurnber 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.

关 键 词：麋鹿 皮肤成纤维细胞 体外培养 核型分析 冷冻保存 

分 类 号：Q2-33[生物学—细胞生物学]