不同血清浓度及介质对体外培养小鼠成纤维细胞L929细胞系生长曲线和细胞形态的影响

The Influence of Different Blood Serum Concentration and Medium to the Growth Curve and the Morphological Chara-cter of Invitro Culture L929 Cell Line of Mouse Fibroblasts

目的:通过探讨不同血清浓度及介质对体外培养小鼠成纤维细胞L929细胞系生长曲线和细胞形态的影响,了解它们对成纤维细胞体外生存和生长的情况。方法:采用体外细胞培养技术进行小鼠成纤维细胞L929细胞,并采用磺基罗丹明B(sulforhodamine B,SRB)酶标仪法(A490nm波长下)测量各孔的吸光值(A值)并转换为小鼠成纤维细胞L929细胞系生长曲线;同时,分别采用HE(苏木精-伊红)染色法、吖啶橙荧光染色和Hoechst33342荧光染色法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细胞系不同浓度的血清分别在3种不同的介质(RPMI1640培养基、0.9%生理盐水和5%葡萄糖生理盐水)中表现出不同的生长和生存情况及细胞形态学改变。其中,在不同血清浓度(0%、10%、20%、30%、40%、50%、60%、70%、80%、90%、100%)的同一介质中,血清浓度为0%和100%浓度组的小鼠成纤维细胞L929细胞生长明昆抑制及吸发光度(A)值呈逐步减少的趋势,而除此之外的其它血清浓度组对小鼠成纤维细胞L929细胞的影响,基本表现为随着血清浓度的升高,吸光度(A)值呈逐步增加及促进细胞生长的趋势。而某些相同血清浓度在不同介质中对小鼠成纤维细胞L929细胞系生长和生存及形态学的有利影响,则基本表现为RPMI1640培养基作为介质的细胞培养效果优于0.9%生理盐水,而0.9%生理盐水作为介质的细胞培养效果又优于5%葡萄糖生理盐水。结论:不同血清浓度及介质对体外培养小鼠成纤维细胞L929细胞系生长和生存及细胞学形态均有明显影响。提示在进行体外细胞培养时应充分考虑细胞培养介质及所添加血清的浓度。

OBJECTIVES： To investigate the influence of different blood serum concentration and medium to the growth curve and the morphological character of invitro culture L929 cell Line of mouse fibroblasts, and to learne the ways how the fibroblasts exist and grow in vitro. METHODS ： The technology of cell invitro culture to the L929 cell line of the mouse fibroblasts were used, and the sulforhodamine B and microplate reader （with a wavelength of under 490nm） were also performed to measure absorbency of every hole, and transformed it into the growth curve of L929 cell line of the fibroblasts. At the same time, The staining methods of HE, acridine orange fluorescence and Hoechst33342 fluorescence were used to observed the morphology of L929 cell line of the mouse fibroblasts. RESULTS： The blood serum of the L929 cell line of the mouse fibroblasts in different concentrations expressed differently in growth rates, survival rates and cell morphology in three different media （ namely RPMI 1640 culture medium, 0.9% normal saline and 5% glucose normal saline）. Different concentrations of blood serum in the same media examined included 0% , 10% , 20%, 30% , 40%, 50%, 60%, 70%, 80%, 90% and 100%. The growth of the L929 cell line of the mouse fibroblasts cultured in 0% and 100% blood serum concentration were apparently restrained in the growth and the absorbency decreased gradually. Besides, in the other concentrations, the absorbency increased and the cell growth was promoted as concentrations increased. As for the same concentration, the RPMI1640 culture medium showed a better effect than the 0. 9% normal saline, which in turn performed better effect than the 5% glucose normal saline. CONCLUSIONS： The different blood serum concentration and medium have significant effects in the growth, survival and morphology of the L929 cell line of the mouse fibroblasts under an invitro condition. The cell culture medium and the blood serum concentration should be fully taken into consideration when the cell is cultured in an invitro condition.