一种鸡胚成纤维细胞快速制备与保存方法的研究

A Study on A Rapid Preparation and Preservation Method of Chicken Embryo Fibroblasts

为了进一步提高鸡胚成纤维细胞(Chicken embryo fibroblast,CEF)的制备效率和活力,试验以短时多次消化法缩短胰酶消化时间,提高细胞活力。同时,通过优化血清、胰酶、培养基等保存条件,增加细胞4℃保存时间。经CCK-8细胞活性检测、活细胞计数、传代培养等方法比较验证,结果表明,所建立的制备方法简易、快速、细胞活力高;细胞在4℃、不添加血清和胰酶的DMEM培养基中保存96 h内仍保有良好的贴壁生长能力。该方法的建立为实验室内快速制备CEF细胞提供候选方案,可有效减少鸡胚的使用。

In order to further improve the preparation efficiency and viability of chicken embryo fibroblast(CEF),the experiment was carried out to shorten the trypsin digestion time by short multiple digestion method to improve the cell viability.At the same time,by optimizing the preservation conditions of serum,trypsin,and culture medium,we grow the time of cell preservation at 4℃.Comparative validation by CCK-8 cell activity assay,viable cell counting,and passaging culture showed that the established preparation method is simple,rapid,and has high cell viability;Cells retained good adherence to the wall during 96 h of storage at 4°C in DMEM medium,no added serum,and no added trypsin.The establishment of this method provides a candidate for rapid preparation of CEF cells in the laboratory,which can effectively reduce the use of chicken embryos.