牛原代肌肉卫星细胞和前体脂肪细胞不同混合比例共培养细胞活性的比较

为探索不同细胞混合比例对牛原代肌肉卫星细胞和前体脂肪细胞共培养体系细胞活性的影响,本研究采集新生秦川犊牛背最长肌和肾周脂肪,分离提取肌卫星细胞和前体脂肪细胞,采用DMEM/F12培养基,建立不同血清浓度(5%、10%、15%、20%的胎牛血清,FBS)的牛原代肌肉卫星细胞和前体脂肪细胞的共培养体系,然后通过分别调整各共培养体系中肌卫星细胞和前体脂肪细胞的混合比例(肌肉细胞∶脂肪细胞=10∶1、5∶1、2∶1),来研究细胞混合比例对各共培养体系的影响。共培养14d,每两天更换培养基,并采用MTT染色法测定细胞活性。统计分析后发现∶随着共培养体系中前体脂肪细胞比例增加,共培养细胞活性增加;共培养10d以后不同混合比例细胞活性有着显著差异(P<0.05);混合比例为2∶1时共培养细胞活性最高。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,建议牛肌卫星细胞和前体脂肪细胞按5∶1或2∶1的混合比例进行共培养。

Comparison of cellular viability under different cell mix proportion co-culture system of bovine primary skeletal muscle satellite cells and preadipocytes

To explore the effect of cellular viability in different cells mixed proportion of co-culture bovine muscle satellite cells and preadipocytes, there were longissimus dorsi and perirenal fat collected from new-born Qinchuan calf to isolate the muscle satellite cells and preadipocytes. Using the common culture medium of DMEM/F12 with different proportions of serum (fetal bovine serum ∶ 5%, 10%, 15%, 20%(v/v)), it was established a co-culture system of 14-days cultivation. In each co-culture system, we changed the cellular mixed ratio (the ratio of SMSC to preadipocytes were 2 ∶1, 5 ∶1, and 10 ∶1), and changed the medium and determined cellular viability by MTT assay in every 2 days, to compare the cellular viability under different mixed proportion. After the statistical analysis, the results showed that, 1) with the increase of the proportion of preadipocytes in this co-culture system, the co-culture cellular viability increased. 2) after 10-day, there was significant difference between different mixed ratio. 3)there was the highest cellular viability at the 2 ∶1 mixing ratio of muscle satellite cells and preadipocytes. So that, in order to obtain the highest cellular viability of co-culture bovine muscle satellite cells and preadipocytes, it was suggested that the mixture ratio were 5 ∶1 or 2 ∶1.