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The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells

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Abstract

Endometrial epithelial cells (EECs) cultured in vitro are valuable tools for investigating embryo implantation and trophoblast differentiation. In this study, we have established the bovine EECs and trophoblast stem-like (TS) coculture system, and used it to investigate the binucleate cell formation of ungulates. The EECs was derived from the uterine horn ipsilateral to the corpus luteum by using collagenase I and deoxyribonuclease I, which exhibited typical epithelial morphology and were expressing bovine uterine epithelial marker such as IFNAR1, IFNAR2, Erα, PGR, ESR1 and KRT18. The cells immunostained positively by epithelial and trophectoderm marker cytokeratin 18 (KRT18) and stromal marker vimentin antibodies, and the KRT18 positive cells reached 99 %. The EECs can be cultured for up to 20 passages in vitro with no significant morphology changes and uterine epithelial marker gene expression alteration. The bTS cells were established in a dual inhibitor system and exhibited typical trophoblast stem cell characteristics. When bTS cells were cultured with EECs, the bTS cells adhered to the EECs as adhering to feeder cells. Binucleate cells began appearing on day 4 of coculture and reached approximately 18.47 % of the differentiated cells. Quantitative real-time PCR or immunofluorescence analyses were performed on bTS cells cocultured at day 6 and day 12. The results showed that the expression level of KRT18 was down-regulated while the expression level of trophoblast differentiation marker MASH2, HAND1, GCM1 and CDX2 was up-regulated in bTS cells. In conclusion, bovine EECs can be obtained from the uterine horn ipsilateral to the corpus luteum via treatment with collagenase I and deoxyribonuclease I, and the EECs-bTS cells coculture system presents an ideal tool for studying the differentiation of bTS cells to trophoblast binucleate cells.

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References

  • Bai H, Sakurai T, Bai R, Yamakoshi S, Aoki E, Kuse M, Okuda K, Imakawa K (2014) Establishment and characterization of immortalized bovine endometrial epithelial cells. Anim Sci J 85:799–804. doi:10.1111/asj.12202

    Article  CAS  Google Scholar 

  • Bergh J, Nilsson K, Dahl D, Andersson L, Virtanen I, Lehto VP (1984) Expression of intermediate filaments in established human lung cancer cell lines. An indicator of differentiation and derivation. Lab Investig J Tech Methods Pathol 51:307–316

    CAS  Google Scholar 

  • Denker HW (1993) Implantation: a cell biological paradox. J Exp Zool 266:541–558. doi:10.1002/jez.1402660606

    Article  CAS  Google Scholar 

  • Fortier MA, Guilbault LA, Grasso F (1988) Specific properties of epithelial and stromal cells from the endometrium of cows. J Reprod Fertil 83:239–248

    Article  CAS  Google Scholar 

  • Hashizume K, Ushizawa K, Patel OV, Kizaki K, Imai K, Yamada O, Nakano H, Takahashi T (2007) Gene expression and maintenance of pregnancy in bovine: roles of trophoblastic binucleate cell-specific molecules. Reprod Fertil Dev 19:79–90. doi:10.1071/Rd06118

    Article  CAS  Google Scholar 

  • Horn S, Bathgate R, Lioutas C, Bracken K, Ivell R (1998) Bovine endometrial epithelial cells as a model system to study oxytocin receptor regulation. Hum Reprod Update 4:605–614

    Article  CAS  Google Scholar 

  • Hou D, Su M, Li X, Li Z, Yun T, Zhao Y, Zhang M, Zhao L, Li R, Yu H, Li X (2015) The efficient derivation of trophoblast cells from porcine in vitro fertilized and parthenogenetic blastocysts and culture with ROCK inhibitor Y-27632. PLoS ONE 10:e0142442. doi:10.1371/journal.pone.0142442

    Article  Google Scholar 

  • Huang X, Han X, Uyunbilig B, Zhang M, Duo S, Zuo Y, Zhao Y, Yun T, Tai D, Wang C, Li J, Li X, Li R (2014) Establishment of bovine trophoblast stem-like cells from in vitro-produced blastocyst-stage embryos using two inhibitors. Stem Cells Dev 23:1501–1514. doi:10.1089/scd.2013.0329

    Article  CAS  Google Scholar 

  • Krishnaswamy N, Danyod G, Chapdelaine P, Fortier MA (2009) Oxytocin receptor down-regulation is not necessary for reducing oxytocin-induced prostaglandin F(2alpha) accumulation by interferon-tau in a bovine endometrial epithelial cell line. Endocrinology 150:897–905. doi:10.1210/en.2008-0704

    Article  CAS  Google Scholar 

  • Li P, Tong C, Mehrian-Shai R, Jia L, Wu N, Yan Y, Maxson RE, Schulze EN, Song H, Hsieh CL, Pera MF, Ying QL (2008) Germline competent embryonic stem cells derived from rat blastocysts. Cell 135:1299–1310. doi:10.1016/j.cell.2008.12.006

    Article  CAS  Google Scholar 

  • Munson L, Ellington JE, Schlafer DH (1991) Bovine trophoblastic cell vesicle attachment to polarized endometrial epithelial cells in vitro. In Vitro Cell Dev Biol 27:31–38

    Article  CAS  Google Scholar 

  • Pagan R, Martin I, Alonso A, Llobera M, Vilaro S (1996) Vimentin filaments follow the preexisting cytokeratin network during epithelial-mesenchymal transition of cultured neonatal rat hepatocytes. Exp Cell Res 222:333–344

    Article  CAS  Google Scholar 

  • Sakurai T, Bai H, Bai R, Arai M, Iwazawa M, Zhang J, Konno T, Godkin JD, Okuda K, Imakawa K (2012) Coculture system that mimics in vivo attachment processes in bovine trophoblast cells. Biol Reprod 87:60. doi:10.1095/biolreprod.112.100180

    Article  Google Scholar 

  • Skarzynski DJ, Miyamoto Y, Okuda K (2000) Production of prostaglandin F2α by cultured bovine endometrial cells in response to tumor necrosis factor α: cell type specificity and intracellular mechanisms. Biol Reprod 62:1116–1120

    Article  CAS  Google Scholar 

  • Thie M, Denker HW (2002) In vitro studies on endometrial adhesiveness for trophoblast: cellular dynamics in uterine epithelial cells. Cells Tissues Organs 172:237–252

    Article  Google Scholar 

  • Wang G, Johnson GA, Spencer TE, Bazer FW (2000) Isolation, immortalization, and initial characterization of uterine cell lines: an in vitro model system for the porcine uterus. In Vitro Cell Dev Biol Anim 36:650–656

    Article  CAS  Google Scholar 

  • Wooding FB, Flint AP (1994) Placentation. In: Lamming GE (ed) Marshall’s physiology of reproduction: volume 3 pregnancy and lactation. Springer, Dordrecht, pp 233–460. doi:10.1007/978-94-011-1286-4_4

    Chapter  Google Scholar 

  • Wooding FB, Morgan G, Monaghan S, Hamon M, Heap RB (1996) Functional specialization in the ruminant placenta: evidence for two populations of fetal binucleate cells of different selective synthetic capacity. Placenta 17:75–86

    Article  CAS  Google Scholar 

  • Yamakoshi S, Bai R, Chaen T, Ideta A, Aoyagi Y, Sakurai T, Konno T, Imakawa K (2012) Expression of mesenchymal-related genes by the bovine trophectoderm following conceptus attachment to the endometrial epithelium. Reproduction 143:377–387. doi:10.1530/REP-11-0364

    Article  CAS  Google Scholar 

  • Zeiler M, Leiser R, Johnson GA, Tinneberg HR, Pfarrer C (2007) Development of an in vitro model for bovine placentation: a comparison of the in vivo and in vitro expression of integrins and components of extracellular matrix in bovine placental cells. Cells Tissues Organs 186:229–242. doi:10.1159/000107947

    Article  CAS  Google Scholar 

  • Zhang S, Lin H, Kong S, Wang S, Wang H, Wang H, Armant DR (2013) Physiological and molecular determinants of embryo implantation. Mol Asp Med 34:939–980. doi:10.1016/j.mam.2012.12.011

    Article  Google Scholar 

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Acknowledgments

This work was supported by National Transgenic Project of China (2014ZX08010-001) and National Natural Sciences Foundation of China (31460309).

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Authors and Affiliations

  1. The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, No. 24 Zhao Jun Rd, Yu Quan Qu, Hohhot, 010070, Inner Mongolia, China

    Xiawei Li, Zhiying Li, Dongxia Hou, Yuhang Zhao, Chen Wang & Xueling Li

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  1. Xiawei Li
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  2. Zhiying Li
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  3. Dongxia Hou
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  4. Yuhang Zhao
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  5. Chen Wang
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  6. Xueling Li
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Correspondence to Xueling Li.

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Li, X., Li, Z., Hou, D. et al. The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells. Cytotechnology 68, 2687–2698 (2016). https://doi.org/10.1007/s10616-016-9994-x

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  • DOI: https://doi.org/10.1007/s10616-016-9994-x

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