无血清条件下UC-MSCs和BMECs共培养对IGF-Ⅰ表达的影响

doi:10.16431/j.cnki.1671-7236.2017.02.016

Abstract

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

CLC Number:

S823

ZHAO Yan-kun, SHAO Wei, LUO Cheng-long, YU Xiong. 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[J]. , 2017, 44(2): 411-416.

References

[1] Fiedler J, Boehm B O, Maisel M,et al. IGF-Ⅰ and IGF-Ⅱstimulate directed cell migration of bone marrow derived human mesenchymal progenitor cells[J].Biochem Biophys Res Commun,2006,24(8):1841-1851.
[2] Ruan W, Kleinberg D L. Insulin-like growth factorⅠis essential for terminal end bud formation and ductal morphogenesis during mammary development[J].Endocrinology,1999,140(11):5078-5081.
[3] 李莹莹,夏小静,吴云娣.胰岛素样生长因子-Ⅰ、肝细胞生长因子、转化生长因子-β1、干扰素-γ对奶牛乳腺上皮细胞增殖活性的影响[J].中国畜牧兽医,2014,41(4):37-42.
[4] Hovey R C, Trott J F, Vonderhaar B K. Establishing a frame work for the functional mammary gland: From endocrinology to morphology[J].J Mammary Gland Biol Neoplasia,2002,7(1):17-38.
[5] Mcbride B W, Barton J L, Gibson J P. Use of recombinant bovine some to tropin for up to two cons ecutive lactation on dairy production traits[J].Dairy Sci,1990,73(11):3448.
[6] 何伯萍,邢艳苹,雷连成,等.IGF-Ⅰ对奶牛乳腺上皮细胞合成乳脂、乳蛋白的影响[J].中国兽医学报,2012,32(8): 1231-1234.
[7] 刘畅,赵锋,李庆章.不同泌乳相关激素和生长因子对奶牛乳腺上皮细胞增殖的影响[J].中国畜牧兽医,2012,39(5):102-106.
[8] Hull K L, Harvey S. Growth hormone and reproduction: A review of endocrine and autocrine/paracrine interactions[J].Int J Endocrinol,2014,29(4):1-24.
[9] Sakamoto K, Yano T, Kobayashi T, et al. Growth hormone suppresses the expression of IGFBP-5,and promotes the IGF-Ⅰinduced phosphorylation of Akt in bovine mammary epithelial cells[J].Domestic Animal Endocrinology,2007,32(4):260-272.
[10] Burgos S, Cant J P.IGF-Ⅰstimulates protein synthesis by enhanced signaling through mTORC1 in bovine mammary epithelial cells[J].Domest Anim Endocrinol, 2010,38(4):211-221.
[11] Rose M T, Weekes T E, Rowlinson P. Correlation of blood and milk components with the milk yield response to bovine somatotropin in dairy cows[J].Domestic Animal Endocrinology,2005,28(3):296-307.
[12] Kleinberg D L, Feldman M, Ruan W.IGF-Ⅰ: An essential factor in terminal end bud formation and ductal morphogenesis[J].J Mammary Gland Biol Neoplasia,2000,10(5):7-19.
[13] Gilmore A P, Valentin J A,Wang P, et al. Activation of BAD by therapeutic inhibition of epidermal growth factor receptor and transactivation by insulin like growth factor receptor[J].J Biol Chem,2002,277(31):27643-27650.
[14] 安培培,郭倩倩,李春义.乳腺发育的研究进展[J].中国畜牧兽医,2015,42(9):2411-2417.
[15] Deeks S, Richards J, Nandi S M. Aintenance of normal rat mammary epithelial cells by insulin and insulin-like growth factorⅠ[J].Exp Cell Res,1988,174(2):448-460.
[16] Kale S, Karihaloo A, Clark P R,et al. Bone marrow stem cells contributes to repair of the ischemically injured renal tubule[J].J Clin Invest,2003,112(5):422-449.
[17] Lincoln D W, Phillips P G, Bove K. Estrogen-induced Ets-1 promotes capillary formation in an in vitro tumor angiogenesis model[J].Breast Cancer Res Treat,2003,78(2):167-178.
[18] Moorehead R A,Fata J E, Johnson M B. Inhibition of mammary epithelial apoptosis and sustained phosphorylation of Akt/PKB in MMTV-IGF-Ⅱtransgenic mice[J]. Cell Death and Differentiation, 2001,8(1):16-29.
[19] Grellier M L,Bordenave L, Amedee J. Cell-to-cell communication between osteogenic and endothelial lineages:Implications for tissue engineering[J].Trends Biotechnology,2009,27(10):562-571.
[20] Kim J W, Kim S Y, Park S Y, et al. Mesenchymal progenitor cells in the human umbilical cord[J]. Ann Hematol, 2004, 83(12):733-738.
[21] 朱孟芝,董雅娟,龚宜超.体外诱导牛骨髓间充质干细胞向乳腺样上皮细胞分化的初步研究[J].中国畜牧兽医, 2012,39(1):92-96.
[22] Sano H I, Narahara S S. Insulin responsiveness to glucose and tissue responsiveness to insulin during lactation in dairy cows[J].Domestic Animal Endocrinol,1993,10(3):191.
[23] Baumrucker C R, Erondu N E. Insulin-like growth factor(IGF) system in the bovine mammary gland and milk[J].Journal of Mammary Gland Biology and Neoplasia,2000,5(1):53-64.
[24] Manjarin R, Steibel J P, Kirkwood R N, et al. Transcript abundance of hormone receptors, mammalian target of rapamycin pathway-related kinases, insulin-like growth factorⅠ, and milk proteins in porcine mammary tissue[J].Animal Science,2012,90(1): 221-230.
[25] 张童非,崔英俊,李庆章.IGF-Ⅰ在奶牛乳腺的表达及其对乳腺上皮细胞泌乳功能的影响[J].中国畜牧兽医, 2012,39(5):126-129.
[26] 龚虹.羊乳腺中IGF-Ⅰ与其受体基因表达量的相关性研究[D].呼和浩特:内蒙古农业大学, 2007.
[27] 崔英俊,李庆章.奶牛不同发育时期乳腺组织IGF-Ⅰ及其受体的表达与定位[J].中国乳品工业,2014,42(6):4-15.
[28] Yang J J, Kennelly J, BarcosV E. The activity of transcription factor Stat5 responds to prolactin, growth hormone, and IGF-Ⅰin rat and bovine mammary explant culture[J].J Anim Sci, 2000, 78(12):3114-3125.

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

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