Culture and Identification of Bovine Endometrial Epithelial Cell

doi:10.11924/j.issn.1000-6850.casb17030211

Abstract

Abstract: The aim of this study was to improve the primary culture method of bovine endometrial cells and to identify their characteristics. Tissue pieces culture method and tissue pieces culture first again after digestion and purification methods were used respectively to isolate and culture bovine endometrial cells. The tissue-specific expression of cytokeratin genes in mammary epithelial cells was identified by immunofluorescent cytochemical staining. The results show that the tissue pieces culture first again after digestion purified method can acquire high percent of live cells and high purity of cells. Therefore, this method is an easy repeatable method for bovine endometrial epithelial cells culture in vitro. The cells obtained by this method can be cultured up to 4-5 passages in vitro, the culture of bovine endometrial cells in vitro establishes a foundation for research of physiological and mechanism of bovine reproduction.

CLC Number:

S814.8

References

[1] Achache H,Revel A. Endometrial receptivity markers,the journey to successful embryo implantation[J].Hum Reprod Update 2006;12:731-746.
[2] Spencer T E,Johnson G A,Bazer F W,et al.Implantation menchanisms:Insight from the sheep[J].Reproduction,2004,128:657-668.
[3] 邝晓娇,张世栋,董书伟,等．奶牛子宫内膜细胞体外培养及应用研究概况[J]. 动物医学进展,2014,35(12):148-152.
[4] Sharkey AM.The endometrium as a cause of implantation failure.Best practice and researcher clinic obs and gyn[J].2003;17(2):289-307.
[5] 王公道,韩旭,黄梦琪,彭丹红. 子宫内膜容受性标志物的研究进展[J]. 东南大学学报(医学版),2015,02:332-336.
[6] 张勇法,杨建英,秦翠丽.小鼠子宫内膜上皮细胞的分离和原代培养.重庆医学[J].2011,40(22)：2236-2237.
[7] 吴庆侠,董海龙,刘忠艳,朱洪云. 永生化山羊子宫内膜上皮细胞系的建立及其生物学特性[J]. 中国农业大学学报,2017,(02):73-79.
[8] 郑欣欣,曹贵方,李琦,李秀男. 绵羊子宫内膜上皮细胞与基质细胞培养方法的建立[J]. 畜牧与兽医,2016,(07):83-88.
[9] 陈伟明,石仙,蔡雯祎,胡嘉嘉,李键. 快速培养纯化牦牛子宫内膜上皮细胞的方法[J]. 黑龙江畜牧兽医,2017,(03):236-239 300.
[10] 都日塔哈拉,曹金山,高龙,付长其,郭羽丽. 奶牛子宫内膜上皮细胞体外培养及鉴定[J]. 中国畜牧兽医,2016,(06):1566-1571.
[11] 李露,史文静,陶爱林,关绮蕙,周龙书. 人子宫内膜细胞原代培养与间质细胞MTS比较[J]. 中国妇幼保健,2017,(01):159-162.
[12] 屠迪.牦牛子宫内膜腺上皮和基质细胞的分离培养和鉴定[D].甘肃农业大学,2006:26-27.
[13] Cao YJ, Zhang CY, Duan EK.The action of fibronectin to development and implantation of mouse blastocysts in vitro[J]. Acta Zoologica Sinica 1999;45:187-193.
[14] Nakayama T, Fujiwara M , Inoue T, Yoshioka S, Mori T, Fuji S.Human peripheral blood mononuclear cells(PBMC)in early pregnancy promote embryo invasion in vitro: HCG enhances the effects of PBMC[J]. Hum Reprod 2002;17:207-212.
[15] Sheldon M ,Cronin J ,Goetze L ,et al.Defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle[J].Biol Reprod ,2009,81:1025-1032.
[16] Herath S ,Deborah P, Dirk W, et al. Expression and funetins of toll-likereceptor 4 in the endometrial cells of the uterus[J].Endocrinol.2006.147(1):562-570.
[17] Srinivasan KR,Dwivedi,Jain SK.Demonstration of uterine receptivity in vitro by co-culture of rat epithelial cells and blastocyst[J].Cell Tissue Res 2006;325:135-141.
[18] Chapwanya A ,Meade K G,Doherty M L,et al.Endomertrial epithelial cells are prtent producers of tracheal antimicrobial peptide and serum amyloid A3 geneexpression in response to E.coli stimulation[J].Vet Immunol Immunopathol,2013,151:157-162.
[19] 何志全,沈文正,窦忠英,等.孕鼠子宫内膜上皮细胞分离、培养及活力检测[J].西北农林科技大学学报:自然科学版,2006,34(2):12-16.
[20] 陈利平,朴学娇,罗永,等.奶牛和绵羊子宫内膜细胞体外培养方法的研究[J].中国农学通报,2012,28(14):99-104.
[21] 赵诚悦,马红,郭镇华,等.猪子宫内膜基质细胞和腺上皮细胞的分离培养[J].吉林农业大学学报,2013,35(4):471-474.
[22] Standerholen F B, Myromslien F D,Kommisrud E, et al.Comparison of electronic volume and forward scater principles of cell selection using fiow cytometry for the evaluation of acrosome and plasma membrane integrity of bull spermatozoa[J].CytometryA.2014,doi:10.1002/cyto.a.22474.