Advertisement

Efficient Production of Murine Uterine Damage Model

  • Yoon Young Kim
  • Bo Bin Choi
  • Ji Won Lim
  • Yong Jin Kim
  • Sung Yob Kim
  • Seung-Yup KuEmail author
Original Article
  • 79 Downloads

Abstract

Background:

Thin or damaged endometrium causes uterine factor-derived infertility resulting in a failure of embryonic implantation. Regeneration of endometrium is a major issue in gynecology and reproductive medicine. Various types of cells and scaffolds were studied to establish an effective therapeutic strategy. For this type of investigations, production of optimal animal models is indispensable. In this study, we tried to establish various murine uterine damage models and compared their features.

Methods:

Three to ten-week-old C57BL/6 female mice were anesthetized using isoflurane. Chemical and mechanical methods using ethanol (EtOH) at 70 or 100% and copper scraper were compared to determine the most efficient condition. Damage of uterine tissue was induced either by vaginal or dorsal surgical approach. After 7–10 days, gross and microscopic morphology, safety and efficiency were compared among the groups.

Results:

Both chemical and mechanical methods resulted in thinner endometrium and reduced number of glands. Gross morphology assessment revealed that the damaged regions of uteri showed various shapes including shrinkage or cystic dilatation of uterine horns. The duration of anesthesia significantly affected recovery after procedure. Uterine damage was most effectively induced by dorsal approach using 100% EtOH treatment compared to mechanical methods.

Conclusion:

Taken together, murine uterine damage models were most successfully established by chemical treatment. This production protocols could be applied further to larger animals such as non-human primate.

Keywords

Uterine damage Endometrium thickness Murine model Embryonic implantation 

Notes

Acknowledgements

This study was supported by the grants of Ministry of Future Planning and Technology and Ministry of Education, Republic of Korea (2016R1E1A1A01943455 and 2016R1D1A1B03934784). The authors appreciate the assistance of Kyu Hyung Park and Amin Tamadon.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Ethical approval

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.
    Kodaman PH, Arici A. Intra-uterine adhesions and fertility outcome: how to optimize success? Curr Opin Obstet Gynecol. 2007;19:207–14.CrossRefGoogle Scholar
  2. 2.
    Yu D, Wong YM, Cheong Y, Xia E, Li TC. Asherman syndrome–one century later. Fertil Steril. 2008;89:759–79.CrossRefGoogle Scholar
  3. 3.
    Tuuli MG, Shanks A, Bernhard L, Odibo AO, Macones GA, Cahill A. Uterine synechiae and pregnancy complications. Obstet Gynecol. 2012;119:810–4.CrossRefGoogle Scholar
  4. 4.
    Lin N, Li X, Song T, Wang J, Meng K, Yang J, et al. The effect of collagen-binding vascular endothelial growth factor on the remodeling of scarred rat uterus following full-thickness injury. Biomaterials. 2012;33:1801–7.CrossRefGoogle Scholar
  5. 5.
    Basir GS, O WS, So WW, Ng EH, Ho PC. Evaluation of cycle-to-cycle variation of endometrial responsiveness using transvaginal sonography in women undergoing assisted reproduction. Ultrasound Obstet Gynecol. 2002;19:484–9.CrossRefGoogle Scholar
  6. 6.
    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
  7. 7.
    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
  8. 8.
    Capella GL, Frigerio E, Fracchiolla C, Altomare G. The simultaneous treatment of inflammatory bowel diseases and associated pyoderma gangrenosum with oral cyclosporin A. Scand J Gastroenterol. 1999;34:220–1.CrossRefGoogle Scholar
  9. 9.
    McWilliams GD, Frattarelli JL. Changes in measured endometrial thickness predict in vitro fertilization success. Fertil Steril. 2007;88:74–81.CrossRefGoogle Scholar
  10. 10.
    Alawadhi F, Du H, Cakmak H, Taylor HS. Bone Marrow-Derived Stem Cell (BMDSC) transplantation improves fertility in a murine model of Asherman’s syndrome. PLoS One. 2014;9:e96662.CrossRefGoogle Scholar
  11. 11.
    Li X, Sun H, Lin N, Hou X, Wang J, Zhou B, et al. Regeneration of uterine horns in rats by collagen scaffolds loaded with collagen-binding human basic fibroblast growth factor. Biomaterials. 2011;32:8172–81.CrossRefGoogle Scholar
  12. 12.
    Jonkman MF, Kauer FM, Nieuwenhuis P, Molenaar I. Segmental uterine horn replacement in the rat using a biodegradable microporous synthetic tube. Artif Organs. 1986;10:475–80.CrossRefGoogle Scholar
  13. 13.
    Taveau JW, Tartaglia M, Buchannan D, Smith B, Koenig G, Thomfohrde K, et al. Regeneration of uterine horn using porcine small intestinal submucosa grafts in rabbits. J Invest Surg. 2004;17:81–92.CrossRefGoogle Scholar
  14. 14.
    Guney M, Oral B, Karahan N, Mungan T. Protective effect of caffeic acid phenethyl ester (CAPE) on fluoride-induced oxidative stress and apoptosis in rat endometrium. Environ Toxicol Pharmacol. 2007;24:86–91.CrossRefGoogle Scholar
  15. 15.
    Guney M, Ozguner F, Oral B, Karahan N, Mungan T. 900 MHz radiofrequency-induced histopathologic changes and oxidative stress in rat endometrium: protection by vitamins E and C. Toxicol Ind Health. 2007;23:411–20.CrossRefGoogle Scholar
  16. 16.
    Hsieh CL, Shiau CS, Lo LM, Hsieh TT, Chang MY. Effectiveness of ultrasound-guided aspiration and sclerotherapy with 95% ethanol for treatment of recurrent ovarian endometriomas. Fertil Steril. 2009;91:2709–13.CrossRefGoogle Scholar
  17. 17.
    Noma J, Yoshida N. Efficacy of ethanol sclerotherapy for ovarian endometriomas. Int J Gynaecol Obstet. 2001;72:35–9.CrossRefGoogle Scholar
  18. 18.
    Moridi I, Mamillapalli R, Cosar E, Ersoy GS, Taylor HS. Bone marrow stem cell chemotactic activity is induced by elevated CXCl12 in endometriosis. Reprod Sci. 2017;24:526–33.CrossRefGoogle Scholar
  19. 19.
    Kim YY, Min H, Kim H, Choi YM, Liu HC, Ku SY. Differential microRNA expression profile of human embryonic stem cell-derived cardiac lineage cells. Tissue Eng Regen Med. 2017;14:163–9.CrossRefGoogle Scholar
  20. 20.
    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
  21. 21.
    Adamczyk L, Adkins JK, Agakishiev G, Aggarwal MM, Ahammed Z, Alekseev I, et al. Measurement of longitudinal spin asymmetries for weak boson production in polarized proton-proton collisions at RHIC. Phys Rev Lett. 2014;113:072301.CrossRefGoogle Scholar
  22. 22.
    Aaby P, Jensen H, Simondon F, Whittle H. High-titer measles vaccination before 9 months of age and increased female mortality: do we have an explanation? Semin Pediatr Infect Dis. 2003;14:220–32.CrossRefGoogle Scholar
  23. 23.
    Schenker JG, Polishuk WZ. Regeneration of rabbit endometrium following intrauterine instillation of chemical agents. Gynecol Invest. 1973;4:1–13.CrossRefGoogle Scholar
  24. 24.
    Milligan SR, Edwards C. The effects of different anaesthetic agents on the estimation of uterine vascular permeability in mice. Lab Anim. 1988;22:343–6.CrossRefGoogle Scholar
  25. 25.
    Matsota P, Kaminioti E, Kostopanagiotou G. Anesthesia related toxic effects on in vitro fertilization outcome: burden of proof. Biomed Res Int. 2015;2015:475362.CrossRefGoogle Scholar
  26. 26.
    Matsota P, Sidiropoulou T, Batistaki C, Giannaris D, Pandazi A, Krepi H, et al. Analgesia with remifentanil versus anesthesia with propofol-alfentanil for transvaginal oocyte retrieval: a randomized trial on their impact on in vitro fertilization outcome. Middle East J Anaesthesiol. 2012;21:685–92.Google Scholar
  27. 27.
    Abraham E, Reinhart K, Opal S, Demeyer I, Doig C, Rodriguez AL, et al. Efficacy and safety of tifacogin (recombinant tissue factor pathway inhibitor) in severe sepsis: a randomized controlled trial. JAMA. 2003;290:238–47.CrossRefGoogle Scholar
  28. 28.
    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
  29. 29.
    Yun JW, Kim YY, Ahn JW, 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
  30. 30.
    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
  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.
    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
  34. 34.
    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
  35. 35.
    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.e1–8.CrossRefGoogle Scholar
  36. 36.
    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
  37. 37.
    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
  38. 38.
    Song T, Zhao X, Sun H, Li X, Lin N, Ding L, et al. Regeneration of uterine horns in rats using collagen scaffolds loaded with human embryonic stem cell-derived endometrium-like cells. Tissue Eng Part A. 2015;21:353–61.CrossRefGoogle Scholar
  39. 39.
    Ding L, Li X, Sun H, Su J, Lin N, Péault B, et al. Transplantation of bone marrow mesenchymal stem cells on collagen scaffolds for the functional regeneration of injured rat uterus. Biomaterials. 2014;35:4888–900.CrossRefGoogle Scholar
  40. 40.
    Miyazaki K, Maruyama T. Partial regeneration and reconstruction of the rat uterus through recellularization of a decellularized uterine matrix. Biomaterials. 2014;35:8791–800.CrossRefGoogle Scholar
  41. 41.
    Zhang XH, Liu ZZ, Tang MX, Zhang YH, Hu L, Liao AH. Morphological changes and expression of cytokine after local endometrial injury in a mouse model. Reprod Sci. 2015;22:1377–86.CrossRefGoogle Scholar
  42. 42.
    Buhimschi CS, Zhao G, Sora N, Madri JA, Buhimschi IA. Myometrial wound healing post-Cesarean delivery in the MRL/MpJ mouse model of uterine scarring. Am J Pathol. 2010;177:197–207.CrossRefGoogle Scholar
  43. 43.
    Micili SC, Goker A, Sayin O, Akokay P, Ergur BU. The effect of lipoic acid on wound healing in a full thickness uterine injury model in rats. J Mol Histol. 2013;44:339–45.CrossRefGoogle Scholar
  44. 44.
    Rinaldi SF, Makieva S, Frew L, Wade J, Thomson AJ, Moran CM, et al. Ultrasound-guided intrauterine injection of lipopolysaccharide as a novel model of preterm birth in the mouse. Am J Pathol. 2015;185:1201–6.CrossRefGoogle Scholar
  45. 45.
    Keskin HL, Sirin YS, Keles H, Turgut O, Ide T, Avsar AF. The aromatase inhibitor letrozole reduces adhesion formation after intraperitoneal surgery in a rat uterine horn model. Eur J Obstet Gynecol Reprod Biol. 2013;167:199–204.CrossRefGoogle Scholar
  46. 46.
    Agostino M, Velkov T, Dingjan T, Williams SJ, Yuriev E, Ramsland PA. The carbohydrate-binding promiscuity of Euonymus europaeus lectin is predicted to involve a single binding site. Glycobiology. 2015;25:101–14.CrossRefGoogle Scholar
  47. 47.
    Zhang Y, Lin X, Dai Y, Hu X, Zhu H, Jiang Y, et al. Endometrial stem cells repair injured endometrium and induce angiogenesis via AKT and ERK pathways. Reproduction. 2016;152:389–402.CrossRefGoogle Scholar
  48. 48.
    Yanpeng W, Qiongxiao H, Sheng XU, Jing S. Establishment of mouse endometrial injury model by curettage or coagulation. Zhejiang Da Xue Xue Bao Yi Xue Ban. 2017;46:186–91.Google Scholar
  49. 49.
    Hamon E, Horvatovich P, Bisch M, Bringel F, Marchioni E, Aoudé-Werner D, et al. Investigation of biomarkers of bile tolerance in Lactobacillus casei using comparative proteomics. J Proteome Res. 2012;11:109–18.CrossRefGoogle Scholar
  50. 50.
    Du H, Naqvi H, Taylor HS. Ischemia/reperfusion injury promotes and granulocyte-colony stimulating factor inhibits migration of bone marrow-derived stem cells to endometrium. Stem Cells Dev. 2012;21:3324–31.CrossRefGoogle Scholar
  51. 51.
    Okazaki M, Matsuyama T, Kohno T, Shindo H, Koji T, Morimoto Y, et al. Induction of epithelial cell apoptosis in the uterus by a mouse uterine ischemia-reperfusion model: possible involvement of tumor necrosis factor-alpha. Biol Reprod. 2005;72:1282–8.CrossRefGoogle Scholar
  52. 52.
    Zhang W, Zhang N, Wang W, Wang F, Gong Y, Jiang H, et al. Efficacy of cefepime, ertapenem and norfloxacin against leptospirosis and for the clearance of pathogens in a hamster model. Microb Pathog. 2014;77:78–83.CrossRefGoogle Scholar
  53. 53.
    Sahin Ersoy G, Zolbin MM, Cosar E, Moridi I, Mamillapalli R, Taylor HS. CXCL12 promotes stem cell recruitment and uterine repair after injury in asherman’s syndrome. Mol Ther Methods Clin Dev. 2017;4:169–77.CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Yoon Young Kim
    • 1
  • Bo Bin Choi
    • 1
  • Ji Won Lim
    • 1
  • Yong Jin Kim
    • 2
  • Sung Yob Kim
    • 3
  • Seung-Yup Ku
    • 1
    Email author
  1. 1.Department of Obstetrics and GynecologySeoul National University College of MedicineSeoulRepublic of Korea
  2. 2.Department of Obstetrics and GynecologyKorea University Guro HospitalSeoulRepublic of Korea
  3. 3.Department of Obstetrics and GynecologyJeju National University School of MedicineJeju-siRepublic of Korea

Personalised recommendations