摘要
目的探讨不同实验条件下不同血清浓度及体外培养时间对小鼠成纤维细胞L929细胞系生长曲线和细胞形态学变化的影响。方法采用体外细胞培养技术进行小鼠成纤维细胞L929细胞的培养,并用磺基罗丹明B（sulforhodamine B,SRB）酶标仪法（A490 nm波长）测量各孔的吸光值（OD值）并转化为小鼠成纤维细胞L929细胞系生长曲线。同时,分别采用HE染色法、吖啶橙荧光染色进行小鼠成纤维细胞L929细胞系形态学观察。结果小鼠成纤维细胞L929细胞系分别在不同浓度的血清中培养2、4、6、8 d,其表现出不同的生长和生存情况及细胞形态学改变。将该细胞系处于相同培养时间和不同血清浓度（0%、20%、40%、60%、80%、100%）时,在20%和40%血清浓度组中,成纤维细胞的生长基本表现为OD值呈现快速增加趋势、且对细胞生长促进作用较强;而血清浓度为0%和100%时对小鼠成纤维细胞L929细胞生长表现出明显抑制作用及OD值呈逐渐减小的趋势。当该细胞系处于不同培养时间、且相同血清浓度时,除0%组OD值呈现减弱趋势,其他浓度组随时间的延长OD值呈增加趋势,在4 d之内OD值增长较快,4~6 d增长趋势减弱,6~8 d增长趋势回升,尤其是100%组。在这种实验条件下,小鼠成纤维细胞L929细胞的形态学也发生了相应改变。结论不同血清浓度及不同培养时间等体外培养环境对小鼠成纤维细胞L929细胞系生长和生存及细胞学形态均有明显影响,提示利用血清进行体外细胞培养时应充分考虑所添加血清的浓度以及细胞培养的时间。
Objective To study the influence of different blood serum concentration and in vitro growth time on the curve and morphological character of in vitro cultured L929 cell line of mouse fibroblasts.Methods L929 cell line of the mouse fibroblasts were cultured by in vitro cell culture technology,and the sulforhodamine B and microplate reader（with a wavelength of under 490 nm） were also performed to measure absorbency of every hole and transform it into growth curve of L929 cell line of the fibroblasts.Meanwhile,staining methods of HE,acridine orange fluorescence and Hoechst33342 fluorescence were respectively used to observe the morphology of L929 cell line of mouse fibroblasts.Results L929 cell line of the mouse fibroblasts,when cultured in blood serum of different concentrations for different time（2,4,6,8 d）,displayed different growth rates,survival rates and cell morphology.When cultured for the same time in blood serum of different concentrations（0%,20%,40%,60%,80%,and 100%）,the results were as follows： when the concentration were 20% and 40%,OD values were tended to increase rapidly and the cell growth were promoted strongly;when the concentrations were 0% and 100%,OD values were tended to decrease steadily and the cell growth was inhibited obviously.When cultured in the same serum for different time,OD values in other groups were tended to increase with time except 0% group.OD values were increased rapidly within the first 4 days,then the increase was slowed down from 4 to 6 days and the tendency of increase was recovered from 6 to 8 days,particularly for 100% group.In this case,morphology of the mouse fibroblasts L929 cells was also changed.Conclusions Different serum concentrations and different growth time in vitro affect cell growth,survival and morphology of mouse L929 fibroblast cell line significantly.Growth time and the blood serum concentration should be fully considered when the cell is cultured in an in vitro condition.This finding provides some reference value for future cytological studies in skin anti-aging.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2011年第11期2020-2024,共5页
Chinese Journal of Gerontology
