不同浓度奶牛乳腺上皮细胞和奶牛脐带间充质干细胞共培养对几种细胞因子分泌的影响研究

Study on the effects of different concentrations of dairy cow mammary epithelial cells co-cultured with dairy cow umbilical cord mesenchymal stem cells on the secretion of several cytokines

为了研究在不同浓度下奶牛脐带间充质干细胞(UC-MSCs)和奶牛乳腺上皮细胞(BMECs)共培养对共培养体系细胞增殖和细胞贴壁率的影响,并通过检测与细胞增殖有关的细胞因子来探究其作用机制,试验将P3代UC-MSCs与P3代BMECs按照浓度比例为1∶1、1∶2、1∶3、1∶4、1∶5、1∶10、1∶50、1∶100、1∶1 000、2∶1等不同比例随机混合培养,同时设立UC-MSCs与BMECs单纯培养组为对照组,并分别于0,24,48,72,96,120和144小时提取上清液检测表皮生长因子(EGF)、碱性成纤维生长因子(b FGF)、转化生长因子α(TGF-α)、转化生长因子β(TGF-β)、肝细胞生长因子(HGF)的分泌水平。结果表明:将UC-MSCs和BMECs按照不同浓度比混合共培养后,EGF、HGF、b FGF分泌水平在72小时时最高,且1∶2浓度组显著高于对照组(P<0.05);TGF-α、TGF-β分泌水平在72小时时最低,且1∶2浓度组显著低于对照组(P<0.05)。说明UC-MSCs和BMECs共培养能提高EGF、HGF、b FGF分泌水平,降低TGF-α、TGF-β的分泌水平。

To study the effects of different concentrations of dairy cow umbilical cord mesenchymal stem cells( UC- MSCs) co- cultured with dairy cow mammary epithelial cells( DCMECs) on cell proliferation and cell adherence rate in the co- culture system,and explore the mechanism of action by detecting cell proliferation- related cytokines. The P3 generation of UC- MSCs and P3 generation of DCMECs were randomly mixed for culture according to different ratios of concentrations,including 1∶1,1∶2,1∶3,1∶4,1 ∶5,1 ∶10,1 ∶50,1 ∶100,1 ∶1 000,2 ∶1 etc.meanwhile,the pure- culture groups for UC- MSCs and DCMECs were established as the control group. The supernatants were extracted to detect the secretion levels of epidermal growth factor( EGF),basic fibroblast growth factor( b FGF),transforming growth factor α( TGF- α),transforming growth factor β( TGF- β),and hepatocyte growth factor( HGF) at 0 h,24 h,48 h,72 h,96 h,120 h and 144 h,respectively. The results showed that the secretion levels of EGF,HGF,and b FGF were highest at 72 h after the UC- MSCs and DCMECs were co- cultured with different concentrations according to different ratios of concentrations,and the group with a concentration of 1 ∶ 2 was significantly higher( P < 0. 05) than the control group; the secretion levels of TGF- α and TGF- β were lowest at 72 h,and the group with a concentration of 1∶2 was significantly lower( P < 0. 05) than the control group. The results indicate that the UC- MSCs co- cultured with DCMECs can improve the secretion levels of EGF,HGF,and b FGF,and reduce the secretion levels of TGF- α and TGF- β.