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寿光黑鸡成纤维细胞的体外培养与冷冻保存
引用本文:王娟,于媛,王跃嗣,马云,焦飞.寿光黑鸡成纤维细胞的体外培养与冷冻保存[J].安徽农业科学,2009,37(6).
作者姓名:王娟  于媛  王跃嗣  马云  焦飞
作者单位:1. 滨州医学院药学院细胞工程教研室,山东烟台,264003
2. 滨州医学院基础学院生物化学教研室,山东烟台,264003
3. 滨州医学院基础学院病理教研室,山东烟台,264003
基金项目:国家自然科学基金,山东省教育厅基金 
摘    要:目的]建立鸡成纤维细胞的体外培养体系。方法]用组织块法和消化法分别分离培养鸡皮肤成纤维细胞;比较细胞生长速率及冻存复苏率。结果]酶消化培养的成纤维细胞原代生长较快,约5 d便可形成单层;2种方法传代细胞的生长速度相似,仅需2~3 d就可形成单层;使用酶消化法和反复贴壁法,经3~4代后,可获得纯化的成纤维细胞;成纤维细胞经冷冻复苏后有75%~80%存活;分离纯化的胚胎和皮肤成纤维细胞的生长曲线均正常,可传至12代,传代后染色体数目不变。结论]采用组织块法及酶消化法培养鸡成纤维细胞均可获得ES细胞建系所需的饲养层细胞。该研究为ES细胞系的成功建立奠定了基础。

关 键 词:皮肤  胚胎  成纤维细胞  体外培养  冷冻保存

Culture in vitro and Cryopreservation of Shouguang Black Chicken Fibroblasts
WANG Juan et al.Culture in vitro and Cryopreservation of Shouguang Black Chicken Fibroblasts[J].Journal of Anhui Agricultural Sciences,2009,37(6).
Authors:WANG Juan
Institution:WANG Juan et al(School of Pharmaceutical Sciences,Binzhou Medical University,Yantai,Sh,ong 264003)
Abstract:Objective] The aim of this study was to establish the in vitro culture system of chicken fibroblasts.Method] Tissue explant method and enzymatic digestion method were used to separate and culture chicken fibroblasts respectively,and the cell growth rates and the cryopreservation and recovery rates were compared.Result] The primary chicken fibroblasts derived from enzymatic digestion grew faster than those derived from tissue explant culture and formed confluent monolayer after culture for 5 days.However,...
Keywords:Skin  Embryonic  Fibroblasts  Culture in vitro  Cryopreservation  
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