腾冲马成纤维细胞系建立及其生物学特性分析  

Establishment of Tengchong Horse Fibroblast Cell Line and Analysis of Its Biological Characteristics

作　　者：付晶 相德才 刘艺端 穆德孟 浦仕奇 吴国权 FU Jing;XIANG Decai;LIU Yiduan;MU Demeng;PU Shiqi;WU Guoquan(National Regional(Yunnan)Gene Bank of Livestock and Poultry Genetic Resources,Yunnan Academy of Animal Husbandry and Veterinary Sciences,Kunming 650224,China;Yunnan Provincial Animal Husbandry Station,Kunming 650224,China)

机构地区：[1]云南省畜牧兽医科学院,国家区域性(云南)畜禽遗传资源基因库,昆明650224 [2]云南省畜牧总站,昆明650224

出　　处：《中国草食动物科学》2025年第2期1-7,共7页China Herbivore Science

基　　金：国家重点研发计划(2024YFD1200702);院士(专家)工作站项目(202305AF150168);国家自然科学基金项目(32260837)。

摘　　要：为建立快速稳定的腾冲马成纤维细胞分离培养体系,采用组织块贴壁培养法和差速胰酶消化法对腾冲马耳组织成纤维原代细胞进行了分离纯化及传代培养,并进行相关生物学特性检测及鉴定。结果表明,腾冲马耳成纤维细胞呈典型的纺锤形、不规则三角形或者多角形漩涡状;细胞冻存前与复苏后存活率均在95%以上;细胞生长态势良好,生长曲线呈“S”型;染色体条数为2n=64,其中31对常染色体,1对性染色体;荧光定量检测到成纤维细胞特征Vimentin基因在2~7代细胞中均有表达,细胞免疫荧光检测波形蛋白(Vimentin)的表达比例达99%;培养的细胞未受微生物污染。综上,试验成功建立了遗传特性稳定的腾冲马成纤维细胞系。In order to establish a rapid and stable isolation and culture system of Tengchong horse fibroblasts,the primary cells of Tengchong horse fibroblasts were isolated,purified and subcultured by tissue block monolayer culture method and differential trypsin digestion method,and the related biological characteristics were detected and identified.The results showed that Tengchong horse ear fibroblasts were typical spindle-shaped,irregular triangle or polygonal vortex;the survival rate of cells before cryopreservation and after resuscitation was above 95%.The growth trend of cells was good,and the growth curve was"s";the number of chromosomes is 2n=64,including 31 pairs of autosomes and 1 pair of sex chromosomes.Fluorescence quantitative detection of fibroblast characteristics Vimentin gene was expressed in 2-7 generations of cells,and the expression ratio of Vimentin was 99%by cellular immunofluorescence detection.The cultured cells were not contaminated by microorganisms.In summary,it shows that the Tengchong horse fibroblast cell line with stable genetic characteristics has been successfully established.

关 键 词：腾冲马 成纤维细胞 分离培养 生物学特性 冷冻保存 

分 类 号：S821.2[农业科学—畜牧学]