血清浓度对共培养条件下牛原代肌肉卫星细胞及前体脂肪细胞活性影响

为探索添加血清浓度对共培养条件下细胞活性的影响,本研究采集新生秦川犊牛背最长肌和肾周脂肪,分离提取前体脂肪细胞和肌卫星细胞,建立以DMEM/F12培养基,不同细胞混合比例(肌肉细胞:脂肪细胞=10:1、5:1、2:1)的多种共培养体系,通过调整各共培养体系培养基中的胎牛血清比例(5%、10%、15%、20%的胎牛血清,FBS),来研究血清浓度对各共培养体系中细胞活性的影响。共培养14d,每两天更换对应培养基,并采用MTT染色法测定共培养细胞的细胞活性。统计分析后发现:共培养细胞活性随着血清浓度的上升而增加;15%FBS和20%FBS浓度下细胞活性均显著高于5%FBS组和10%FBS组(P<0.05);虽20%FBS组细胞活性高于15%组,但差异不显著(P>0.05)。综上,为了获得最好的牛肌卫星细胞和前体脂肪细胞共培养效果,达到较高的细胞培养活性,建议采用15%(v/v)以上的FBS进行共培养。

Effect of Serum Concentration on Cellular Viability in Co-Culture System of Bovine Primary Skeletal Muscle Satellite Cells and Preadipocytes

To explory the effect of ditfeant serum concentration on the cellulaa /ability of co - cultura bovine muscle satellite cells and prcadipocytesin, longissimus dorsiand perirenal fat were collected from new 一 bom Qinchuan celf fo isolate ehe muscle satellite cells and preadipocytes. Using the common cultury medium of DMEM/F12 and different mixed ratin (the ratin of SMSC te preadipocytes wery 2- 1, 5- 1, and 10: 1), it was established co-cultury systems of 14-days cultivation. In each co - cultura system, we adjusted the cultura medium with dyferent propoAions of serum fetal bovine serum: 5%, 10%, 15%, 20%(v/v), te explory the efed of serum conontration on these co-cultura systems. At evey 2 days, we changed the medium and determined cellular viability by MTT asay. After tUe statisticel analysis, the results showed that the cellular /dbility increesed with the increese of serum conontration. The celluIco viability which cultuad with 15% FBS or 20% FBS was sinnificently higher tUan tUat cultured with 5% FBS or 10% FBS ( P < 0. 05 ), respectively. Although the cellular viability of 20% FBS group was higher than the group of 15% FBS, there was no significent dUferenco(P>0.05 ). So that, in order te obtain the best effed of co 一 culture bovine muscle satellite cells wit preadipocytes, it was suggested that Uic coculture medium should be the DMEM/F12 with the FBS more than 15%( v/v).