《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (2): 411-416.doi: 10.16431/j.cnki.1671-7236.2017.02.016

• 生理生化 • 上一篇    下一篇

无血清条件下UC-MSCs和BMECs共培养对IGF-Ⅰ表达的影响

赵艳坤, 邵伟, 雒诚龙, 余雄   

  1. 新疆农业大学动物科学学院, 新疆肉乳用草食动物营养实验室, 乌鲁木齐 830052
  • 收稿日期:2016-08-23 出版日期:2017-02-20 发布日期:2017-02-25
  • 通讯作者: 余雄 E-mail:yuxiong8763601@126.com
  • 作者简介:赵艳坤(1990-), 女, 河南郑州人, 博士生, 研究方向:动物营养与饲料科学, E-mail:452349621@qq.com
  • 基金资助:

    奶产业体系资助(cars-37);2015年国家自然科学基金(31560645);中国博士后基金委;新疆维吾尔自治区高等学校科研计划项目资助(XJEDU2013S17)

Effect of Umbilical Cord Mesenchymal Stem Cells and Bovine Mammary Gland Epithelial Cells Co-culture under the Serum-free Condition on Expression of IGF-Ⅰ

ZHAO Yan-kun, SHAO Wei, LUO Cheng-long, YU Xiong   

  1. Xinjiang Meat Emulsions with Plant-eating Animal Nutrition Laboratory, College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China
  • Received:2016-08-23 Online:2017-02-20 Published:2017-02-25

摘要:

试验旨在探讨无血清条件下脐带间充质干细胞(UC-MSCs)和奶牛乳腺上皮细胞(BMECs)共培养对类胰岛素样生长因子-Ⅰ(IGF-Ⅰ)表达的影响,将UC-MSCs和BMECs按1:2直接共培养,对比单纯培养的两种细胞,48 h时利用ELISA法检测各组上清IGF-Ⅰ浓度水平,再利用Transwell小室将两种细胞分离培养,实时荧光定量 PCR检测各组细胞IGF-ⅠR、IGF-Ⅰ mRNA的表达。结果显示,共培养中IGF-Ⅰ浓度显著高于UC-MSCs组(P < 0.05),极显著高于BMECs组(P < 0.01);UC-MSCs/BMECs、BMECs/UC-MSCs组IGF-Ⅰ mRNA表达值均极显著高于单纯培养组(P < 0.01),BMECs/UC-MSCs组显著高于UC-MSCs/BMECs组(P < 0.05);UC-MSCs/BMECs、BMECs/UC-MSCs组IGF-ⅠR mRNA表达值显著或极显著高于单纯培养组(P < 0.05;P < 0.01),BMECs/UC-MSCs组较UC-MSCs/BMECs组差异显著(P < 0.05)。综上,体外无血清培养条件下,UC-MSCs和BMECs共培养可显著提高IGF-ⅠR及IGF-Ⅰ mRNA表达,IGF-Ⅰ浓度水平和IGF-ⅠR mRNA的表达具有一致性,IGF-Ⅰ主要存在于UC-MSCs中。

关键词: 脐带间充质干细胞; 奶牛乳腺上皮细胞; 无血清; 共培养; 类胰岛素样生长因子-Ⅰ; 实时荧光定量PCR

Abstract:

The study was aimed to explore the effect of umbilical cord mesenchymal stem cells(UC-MSCs) and bovine mammary gland epithelial cells (BMECs) under the serum-free co-culturing condition on expression of IGF-Ⅰ. UC-MSCs and BMECs were co-cultured directly at the concentration ratios of 1:2,in control groups, UC-MSCs and BMECs were cultured alone.Using ELISA method to detect the IGF-Ⅰlevels in each group supernatant at 48 h, and two kinds of cells were separated by Transwell Chambers, the IGF-Ⅰ and IGF-ⅠR mRNA expression values of each group were estimated with Real-time PCR. The results showed that the IGF-Ⅰ concentration of UC-MSCs and BMECs mixed co-culture was significantly higher than the UC-MSCs group (P < 0.05),and extremely significantly higher than the BMECs group (P < 0.01); The IGF-Ⅰ mRNA expression of UC-MSCs/BMECs and BMECs/UC-MSCs groups were extremely significantly higher than the control group (P < 0.01),and the IGF-Ⅰ mRNA expression of BMECs/UC-MSCs group was significantly higher than the UC-MSCs/BMECs group (P < 0.05); The IGF-ⅠR mRNA expression of UC-MSCs/BMECs and BMECs/UC-MSCs groups were higher than the control group (P < 0.05;P < 0.01), BMECs/UC-MSCs group had significant difference compared with UC-MSCs/BMECs group (P < 0.05). Conclusively, the co-culture of UC-MSCs and BMECs was able to improve the IGF-ⅠR and IGF-Ⅰ mRNA expression under the serum-free condition in vitro,and the IGF-Ⅰ concentration level was correspondence with the IGF-ⅠR mRNA expression, IGF-Ⅰmainly existed in UC-MSCs.

Key words: umbilical cord mesenchymal stem cells (UC-MSCs); bovine mammary epithelial cells(BMECs); serum free; co-culture; IGF-Ⅰ; Real-time PCR

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