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ICR胎鼠成纤维细胞培养方法的优化
引用本文:王英,刘石磊,刘振伟,刘云海,倪和民,邓桂馨,郭勇.ICR胎鼠成纤维细胞培养方法的优化[J].中国实验动物学杂志,2012(8):48-50,55,F0004.
作者姓名:王英  刘石磊  刘振伟  刘云海  倪和民  邓桂馨  郭勇
作者单位:北京农学院动物科学技术学院,北京102206
基金项目:国家自然基金面上资助项目(31072185,30871836),家禽产业技术体系北京市创新团队项目(2011,饲料营养功能).
摘    要:目的确定胎鼠成纤维细胞(mouse embryonic fibroblasts,MEF)最优制备方法,为MEF的制备节省时间与原材料。方法取胎鼠组织,采用组织块培养法、胰酶消化培养法、胰酶消后组织块培养法进行分离培养MEF,比较观察MEF纯度、生长速度、细胞形态等指标,筛选最节省时间与原料的一种培养方法。结果组织块培养法所得MEF生长速度慢,细胞体积小,杂细胞少,纯化所需传代次数少。胰酶消化培养法的MEF生长速度快,体积较大,杂细胞数量多,纯化所需传代次数较多。胰酶消化后组织块培养法的消化后组织块的成纤维细胞生长速度介于前两种方法之间,细胞体积和形态与组织块法培养结果相似,杂细胞数量较少,纯化所需传代次数少。结论胰酶消化结合组织块培养法能最大限度的利用材料,在短时间内可制备大量MEF,满足核移植试验与作为饲养层细胞的需求。因此,胰酶消化结合组织块培养法是MEF最优培养方法。

关 键 词:ICR小鼠  胚胎成纤维细胞  培养

Optimization of the Culture Method of ICR Mouse Embryonic Fibroblasts
WANG Ying,LIU Shi-lei,LIU Zhen,LIU Yun-hai,NI He-min,DENG Gui-xin,GUO Yong.Optimization of the Culture Method of ICR Mouse Embryonic Fibroblasts[J].Chinese Journal of Laboratory Animal Science,2012(8):48-50,55,F0004.
Authors:WANG Ying  LIU Shi-lei  LIU Zhen  LIU Yun-hai  NI He-min  DENG Gui-xin  GUO Yong
Institution:(College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China)
Abstract:Objective To investigate a most suitable method for harvesting mouse embryonic fibroblasts (MEF) ). Methods Three different protocols including tissue only, cell-trypsin digestion, and tissue-trypsin digestion were applied for harvesting and culturing MEF. The MEF growth speed and cell morphology were also compared and analyzed respectively among the 3 methods. Results The cell proliferation was slow, cell size was small, but cells of other types were very few in the tissue only method, but the duration of purification was short. In the cell-trypsin digestion method, the cell proliferation was fast, cell size was somewhat large, but there were a little bit more cells of other types and the duration of purification was somehow longer. Finally, in the tissue-trypsin digestion method, the cell proliferation was very slow, cell size was very small, but there were also very few cells of other types. The duration of purification was very short. Conclusions Our findings indicate that the most efficient method for collecting and culturing mouse embryonic fibroblasts is the protocol of tissue-trypsin digestion.
Keywords:ICR mouse  Embryonic fibroblast  Culture
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