鸭胚成纤维细胞培养、传代及保存方法的研究  

Culture,Subculture and Cryopreservation of Embryo Fibroblast of Duck

作　　者：许静[1,2] 李进军[1] 熊胜[1] 陈黎[1] 杨倩[2] 卢立志[1] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,浙江杭州310021 [2]南京农业大学动物医学院,江苏南京210095

出　　处：《中国家禽》2012年第13期9-13,共5页China Poultry

基　　金：现代农业产业技术体系建设专项资金(NYCYTX-45-02);浙江省农科院科技创新能力提升工程

摘　　要：本研究旨在建立鸭胚成纤维细胞离体培养、传代以及冷冻保存体系,为鸭胚胎或生殖干细胞的离体培养奠定基础。利用胰酶-EDTA消化鸭胚胎组织,10% FBS+DEME营养液培养,获得原代和传代成纤维细胞。分别用DMSO、甘油以及乙二醇作为冷冻保护剂对F1代鸭胚成纤维细胞进行冷冻保存,检测复苏后细胞的生长情况。利用此方法可以获得高活性的鸭胚成纤维细胞,并可成功传至第三代。其中F1代与F2代细胞活力最好,达到90%以上。3种冷冻剂保存的细胞复苏后均与F1代细胞的活力存在明显差异,均小于80%,其中用DMSO冷冻保存的细胞复苏后活力最强,生长密度保持较好。Effective methods for isolation,culture,subculture and cryopreservation of duck embryo fibroblast in vitro was established which would lay the foundation for culture of embryo or germ stem cells. The duck embryonal tissue was digested with trypsin-EDTA to obtain primary cells and passage cells,which were cultured with 10%FBS＋DEME. Use the DMSO, glycerin, ethylene glycol as cryoprotectants to freeze FI cells, and observe the cell viability of anabiotic cells. The methods could be used for isolation and pure culture of duck embryo fibroblast in vitro, and could be subcultured to the third generation. The results indicated that the cell viability in F1 and F2 came up to 90%,but no significant difference was observed. The cell viability of all anabiotic cells decreased （〈80%） after cryostoraged by three different cryoprotectants. DMSO was the most effective reagent for cryopreservation of duck embryo fibroblast.

关 键 词：鸭胚成纤维细胞 体外培养 冷冻保存 细胞活力 

分 类 号：S858.32[农业科学—临床兽医学]