不同品种牛血清对3T3L1前脂肪细胞增殖与分化的影响

Effects of Different Species Bovine Sera on Proliferation and Differentiation in 3T3L1 Preadipocyte

研究不同品种牛血清对3T3-L1细胞增殖与分化的影响,采用模型细胞体外培养法模拟牛脂肪组织生长环境,为牛大理石纹肉早期选择提供一种可能性的方法。抽取17头秦川牛和28头秦杂牛血清,先制备没灭活和灭活血清组培养细胞,MTT法检测细胞增殖相对数,油红O检测脂肪含量,再用不同品种牛血清培养细胞,检测细胞增殖相对数和脂肪含量,以及用比色法测定三磷酸甘油脱氢酶(GPDH)和脂肪酸合成酶(FAS)活性。结果表明,灭活血清培养细胞的增殖相对数量在分化培养第2和4天与分化脂肪含量在第2、4、6和8天都极显著高于没灭活血清组(P<0.01);秦杂牛血清培养细胞的数量在第2和4天显著高于秦川牛(P<0.05),秦川牛血清培养细胞分化的脂肪含量在第8天显著高于秦杂牛(P<0.05),其他天数没有显著性差异(P>0.05);分化第8天的细胞内GPDH和FAS酶活两牛种间没有显著性差异(P>0.05)。结果显示自制血清灭活比没灭活的更利于细胞的培养;牛血清品种是影响前脂肪细胞增殖与分化的因素,秦杂牛血清可能更有利于前脂肪细胞的增殖,而秦川牛血清可能更有利于前脂肪细胞的分化,但仍需进一步研究。

The purpose of this study was to know the effects of different species bovine sera on the proliferation and differentiation of 3T3-L1 preadipocytes.In order to provide the possible method of early selection for marbling,we use cell model in vitro culture to simulate the environment of bovine adipose tissue.Collected the bovine sera of different species from seventeen Qinchuan cattle and twenty-eight Qinchuan-Angus hybrid cattle.One part of sera were heat inactivated.The other were activated.The relatively cell number,which stands for proliferation of 3T3-L1 cells,was assayed by MTT colorimetry.Fat content and enzyme activities,which contain glycerol phosphate dehydrogenase（GPDH） and fatty acid synthetase（FAS）,represented the differentiation.Oil red O staining was used to determine the fat content,and enzyme activities were determined by colorimetry after inducting adipogenesis.The results showed as follow.First,heat inactivated sera were quite significantly higher than activated ones on relatively cell number of 3T3-L1 cell at day 2 and 4,and it＇s the same results with fat content at day 2,4,6 and 8（P〈0.01）.Second,Qinchuan-Angus hybrid bovine sera were significantly higher than Qinchuan bovine sera on relatively cell number at day 2 and 4 in cell culture（P〈0.05）.Qinchuan sera were significantly higher than hybrid sera on lipid content at day 8（P〈0.05）.However,there were no significant different at other days on cell number and lipid content（P〉0.05）.Moreover,there were no significantly different between two species of cattle sera on GPDH and FAS activities in cell culture at day 8（P〉0.05）.Self-made bovine sera within heat inactivation promoted proliferation and differentiation of 3T3-L1 cell.Bovine sera is one of the factors that affect proliferation and differentiation of 3T3-L1 cell.Qinchuan-Angus hybrid bovine sera may be good for proliferation,wahile Qinchuan bovine sera may be good for differentiation.More studys are needed.