Advertisement

Proliferation Profile of Uterine Endometrial Stromal Cells during In Vitro Culture with Gonadotropins: Recombinant versus Urinary Follicle Stimulating Hormone

  • Yong Jin Kim
  • Yoon Young Kim
  • Da Young Song
  • Sang Hoon Lee
  • Chan Woo Park
  • Hoon Kim
  • Seung-Yup KuEmail author
Original Article
  • 40 Downloads

Abstract

Background:

Provision of optimal endometrial stromal cells is essential in uterine tissue engineering. Culture of these cells is significantly influenced by gonadotropin hormones. This investigation attempted to define the proliferation profiles of murine uterine endometrial stromal cells during in vitro culture with recombinant follicle stimulating hormone (rFSH), urinary follicle stimulating hormone (uFSH), and human chorionic gonadotropin (hCG).

Methods:

Murine uterine endometrial stromal cells were collected from 8-week-old mice and cultured in vitro up to 72 h, with rFSH, uFSH, or hCG. Cell cycles were analyzed by BrdU assay, and cyclin D1 expression was evaluated according to dose and duration of gonadotropin treatment.

Results:

BrdU assay showed a further inhibitory effect on murine uterine endometrial stromal cell proliferation when cultured with rFSH compared to uFSH, and a similar inhibitory proliferation profile when cultured with hCG at a specific range of concentrations. The expression of cyclin D1 of murine uterine endometrial stromal cells was down-regulated when cultured with rFSH, uFSH, or hCG, compared to control.

Conclusions:

FSH may inhibit the proliferation of murine uterine endometrial stromal cells during in vitro culture. rFSH may have more significant inhibitory effects on the proliferation of endometrial stromal cells than uFSH. Establishing an optimal endocrine milieu is necessary using more advanced combination of female hormones for in vitro culture of this type of cells.

Keywords

Uterus Endometrial stromal cells In vitro culture 

Notes

Acknowledgements

This study was supported by the grants of Ministry of Future Planning and Technology, Republic of Korea (2016R1E1A1A01943455 and 2016R1D1A1A02937287). The authors appreciate Hye Min Kim for her assistance in the early stage work of this project.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interests.

Ethical statement

All of the animal studies were performed after receiving an approval of the Institutional Animal Care and Use Committee of the Biomedical Research Institute at the Seoul National University Hospital (SNUH-IACUC No. 15-0032).

References

  1. 1.
    Evans GE, Phillipson GTM, Sykes PH, McNoe LA, Print CG, Evans JJ. Does the endometrial gene expression of fertile women vary within and between cycles? Hum Reprod. 2018;33:452–63.CrossRefGoogle Scholar
  2. 2.
    Kim YJ, Kim YY, Kim DW, Joo JK, Kim H, Ku SY. Profile of microRNA expression in endometrial cell during in vitro culture according to progesterone concentration. Tissue Eng Regen Med. 2017;14:617–29.CrossRefGoogle Scholar
  3. 3.
    Eissa AM, Barros FSV, Vrljicak P, Brosens JJ, Cameron NR. Enhanced differentiation potential of primary human endometrial cells cultured on 3D scaffolds. Biomacromolecules. 2018;19:3343–50.CrossRefGoogle Scholar
  4. 4.
    Kim YJ, Kim YY, Kang BC, Kim MS, Ko IK, Liu HC, et al. Induction of multiple ovulation via modulation of angiotensin II receptors in in vitro ovarian follicle culture models. J Tissue Eng Regen Med. 2017;11:3100–10.CrossRefGoogle Scholar
  5. 5.
    Kim YJ, Ku SY, Kim YY, Liu HC, Chi SW, Kim SH, et al. MicroRNAs transfected into granulosa cells may regulate oocyte meiotic competence during in vitro maturation of mouse follicles. Hum Reprod. 2013;28:3050–61.CrossRefGoogle Scholar
  6. 6.
    Kim YJ, Ku SY, Rosenwaks Z, Liu HC, Chi SW, Kang JS, et al. MicroRNA expression profiles are altered by gonadotropins and vitamin C status during in vitro follicular growth. Reprod Sci. 2010;17:1081–9.CrossRefGoogle Scholar
  7. 7.
    Kim YY, Yun JW, Kim JM, Park CG, Rosenwaks Z, Liu HC, et al. Gonadotropin ratio affects the in vitro growth of rhesus ovarian preantral follicles. J Investig Med. 2016;64:888–93.CrossRefGoogle Scholar
  8. 8.
    Gilbert SB, Roof AK, Rajendra Kumar T. Mouse models for the analysis of gonadotropin secretion and action. Best Pract Res Clin Endocrinol Metab. 2018;32:219–39.CrossRefGoogle Scholar
  9. 9.
    Colacurci N, Caprio F, La Verde E, Trotta C, Ianniello R, Mele D, et al. Sequential protocol with urinary-FSH/recombinant-FSH versus standard protocol with recombinant-FSH in women of advanced age undergoing IVF. Gynecol Endocrinol. 2014;30:730–3.CrossRefGoogle Scholar
  10. 10.
    Kim YJ, Ku SY, Jee BC, Suh CS, Kim SH, Choi YM, et al. Effects of adding luteinizing hormone activity to gonadotropin releasing hormone antagonist protocols may differ according to age. Gynecol Endocrinol. 2010;26:256–60.CrossRefGoogle Scholar
  11. 11.
    Chen Y, Zhang Y, Hu M, Liu X, Qi H. Timing of human chorionic gonadotropin (hCG) hormone administration in IVF/ICSI protocols using GnRH agonist or antagonists: a systematic review and meta-analysis. Gynecol Endocrinol. 2014;30:431–7.CrossRefGoogle Scholar
  12. 12.
    Choi YS, Ku SY, Jee BC, Suh CS, Choi YM, Kim JG, et al. Comparison of follicular fluid IGF-I, IGF-II, IGFBP-3, IGFBP-4 and PAPP-A concentrations and their ratios between GnRH agonist and GnRH antagonist protocols for controlled ovarian stimulation in IVF-embryo transfer patients. Hum Reprod. 2006;21:2015–21.CrossRefGoogle Scholar
  13. 13.
    La Marca A, Carducci Artenisio A, Stabile G, Rivasi F, Volpe A. Evidence for cycle-dependent expression of follicle-stimulating hormone receptor in human endometrium. Gynecol Endocrinol. 2005;21:303–6.CrossRefGoogle Scholar
  14. 14.
    Zhang D, Li J, Xu G, Zhang R, Zhou C, Qian Y, et al. Follicle-stimulating hormone promotes age-related endometrial atrophy through cross-talk with transforming growth factor beta signal transduction pathway. Aging Cell. 2015;14:284–7.CrossRefGoogle Scholar
  15. 15.
    Zhang M, Shi H, Segaloff DL, Van Voorhis BJ. Expression and localization of luteinizing hormone receptor in the female mouse reproductive tract. Biol Reprod. 2001;64:179–87.CrossRefGoogle Scholar
  16. 16.
    Ku SY, Choi YM, Suh CS, Kim SH, Kim JG, Moon SY, et al. Effect of gonadotropins on human endometrial stromal cell proliferation in vitro. Arch Gynecol Obstet. 2002;266:223–8.CrossRefGoogle Scholar
  17. 17.
    Park KE, Ku SY, Jung KC, Liu HC, Kim YY, Kim YJ, et al. Effects of urinary and recombinant gonadotropins on in vitro maturation outcomes of mouse preantral follicles. Reprod Sci. 2013;20:909–16.CrossRefGoogle Scholar
  18. 18.
    Huh Y, Kim YY, Ku SY. Perspective of bioartificial uterus as gynecological regenerative medicine. Tissue Eng Regen Med. 2012;9:233–9.CrossRefGoogle Scholar
  19. 19.
    Tamadon A, Park KH, Kim YY, Kang BC, Ku SY. Efficient biomaterials for tissue engineering of female reproductive organs. Tissue Eng Regen Med. 2016;13:447–54.CrossRefGoogle Scholar
  20. 20.
    Yun JW, Kim YY, Ahn JH, Kang BC, Ku SY. Use of nonhuman primates for the development of bioengineered female reproductive organs. Tissue Eng Regen Med. 2016;13:323–34.CrossRefGoogle Scholar
  21. 21.
    Kim YJ, Ku SY, Jee BC, Suh CS, Kim SH, Choi YM, et al. A comparative study on the outcomes of in vitro fertilization between women with polycystic ovary syndrome and those with sonographic polycystic ovary-only in GnRH antagonist cycles. Arch Gynecol Obstet. 2010;282:199–205.CrossRefGoogle Scholar
  22. 22.
    Kim JJ, Choi YM, Chae SJ, Hwang KR, Yoon SH, Kim MJ, et al. Vitamin D deficiency in women with polycystic ovary syndrome. Clin Exp Reprod Med. 2014;41:80–5.CrossRefGoogle Scholar
  23. 23.
    Kim SM, Kim SH, Lee JR, Jee BC, Ku SY, Suh CS, et al. Association of leptin receptor polymorphisms Lys109Arg and Gln223Arg with serum leptin profile and bone mineral density in Korean women. Am J Obstet Gynecol. 2008;198:421.CrossRefGoogle Scholar
  24. 24.
    Lee SH, Lee S, Jun HS, Jeong HJ, Cha WT, Cho YS, et al. Expression of the mitochondrial ATPase6 gene and Tfam in down syndrome. Mol Cells. 2003;15:181–5.Google Scholar
  25. 25.
    Kim YY, Tamadon A, Ku SY. Potential use of antiapoptotic proteins and noncoding RNAs for efficient in vitro follicular maturation and ovarian bioengineering. Tissue Eng Part B Rev. 2017;23:142–58.CrossRefGoogle Scholar
  26. 26.
    Chang CC, Hsieh YY, Hsu KH, Lin CS. Effects of gonadotropins (Gonal-F and Puregon) on human endometrial cell proliferation in vitro. Taiwan J Obstet Gynecol. 2011;50:42–7.CrossRefGoogle Scholar
  27. 27.
    Sacchi S, Sena P, Degli Esposti C, Lui J, La Marca A. Evidence for expression and functionality of FSH and LH/hCG receptors in human endometrium. J Assist Reprod Genet. 2018.  https://doi.org/10.1007/s10815-018-1248-8.Google Scholar
  28. 28.
    Evans J, Salamonsen LA. Too much of a good thing? Experimental evidence suggests prolonged exposure to hCG is detrimental to endometrial receptivity. Hum Reprod. 2013;28:1610–9.CrossRefGoogle Scholar
  29. 29.
    Cameo P, Szmidt M, Strakova Z, Mavrogianis P, Sharpe-Timms KL, Fazleabas AT. Decidualization regulates the expression of the endometrial chorionic gonadotropin receptor in the primate. Biol Reprod. 2006;75:681–9.CrossRefGoogle Scholar
  30. 30.
    Sherwin JR, Hastings JM, Jackson KS, Mavrogianis PA, Sharkey AM, Fazleabas AT. The endometrial response to chorionic gonadotropin is blunted in a baboon model of endometriosis. Endocrinology. 2010;151:4982–93.CrossRefGoogle Scholar
  31. 31.
    Kim YY, Kim YJ, Cho KM, Kim SH, Park KE, Kang BC, et al. The expression profile of angiotensin system on thawed murine ovaries. Tissue Eng Regen Med. 2016;13:724–31.CrossRefGoogle Scholar
  32. 32.
    Kim YJ, Park KE, Kim YY, Kim H, Ku SY, Suh CS, et al. Effects of estradiol on the paracrine regulator expression of in vitro maturated murine ovarian follicles. Tissue Eng Regen Med. 2017;14:31–8.CrossRefGoogle Scholar
  33. 33.
    Yang H, Lei CX, Zhang W. Human chorionic gonadotropin (hCG) regulation of galectin-3 expression in endometrial epithelial cells and endometrial stromal cells. Acta Histochem. 2013;115:3–7.CrossRefGoogle Scholar

Copyright information

© The Korean Tissue Engineering and Regenerative Medicine Society and Springer Nature B.V. 2018

Authors and Affiliations

  • Yong Jin Kim
    • 1
  • Yoon Young Kim
    • 2
  • Da Young Song
    • 2
  • Sang Hoon Lee
    • 1
  • Chan Woo Park
    • 3
  • Hoon Kim
    • 2
  • Seung-Yup Ku
    • 2
    Email author
  1. 1.Department of Obstetrics and GynecologyKorea University Medical CenterSeoulRepublic of Korea
  2. 2.Department of Obstetrics and GynecologySeoul National University HospitalSeoulRepublic of Korea
  3. 3.Department of Obstetrics and GynecologyCheil General HospitalSeoulRepublic of Korea

Personalised recommendations