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Home / Equine Research Bank / Int J Mol Sci / Production of Mare Chorionic Girdle Organoids That Secrete E...
International journal of molecular sciences2023; 24(11); 9538; doi: 10.3390/ijms24119538

Production of Mare Chorionic Girdle Organoids That Secrete Equine Chorionic Gonadotropin.

Abstract: The equine chorionic girdle is comprised of specialized invasive trophoblast cells that begin formation approximately 25 days after ovulation (day 0) and invade the endometrium to become endometrial cups. These specialized trophoblast cells transition from uninucleate to differentiated binucleate trophoblast cells that secrete the glycoprotein hormone equine chorionic gonadotropin (eCG; formerly known as pregnant mare serum gonadotropin or PMSG). This eCG has LH-like activity in the horse but variable LH- and FSH-like activity in other species and has been utilized for these properties both in vivo and in vitro. To produce eCG commercially, large volumes of whole blood must be collected from pregnant mares, which negatively impacts equine welfare due to repeated blood collections and the birth of an unwanted foal. Attempts to produce eCG in vitro using long-term culture of chorionic girdle explants have not been successful beyond 180 days, with peak eCG production at 30 days of culture. Organoids are three-dimensional cell clusters that self-organize and can remain genetically and phenotypically stable throughout long-term culture (i.e., months). Human trophoblast organoids have been reported to successfully produce human chorionic gonadotropin (hCG) and proliferate long-term (>1 year). The objective of this study was to evaluate whether organoids derived from equine chorionic girdle maintain physiological functionality. Here we show generation of chorionic girdle organoids for the first time and demonstrate in vitro production of eCG for up to 6 weeks in culture. Therefore, equine chorionic girdle organoids provide a physiologically representative 3D in vitro model for chorionic girdle development of early equine pregnancy.
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Publication Date: 2023-05-31 PubMed ID: 37298490PubMed Central: PMC10253530DOI: 10.3390/ijms24119538Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research focuses on the production of equine chorionic gonadotropin (eCG), a glycoprotein hormone, by utilizing specialized cells from a horse’s chorionic girdle in a 3D organoid model. This new method shows promise as a more sustainable, welfare-conscious way to produce eCG, which has traditionally been sourced from pregnant mares’ blood, causing unwanted implications for animal welfare.

Research Background

  • Equine Chorionic Gonadotropin (eCG) is a hormone produced during early equine pregnancy with important roles in stimulating luteinizing hormone-like activity in the horse and feritilzation.
  • Commercially, eCG is extracted from the blood of pregnant mares, leading to adverse impact on equine welfare due to frequent blood collections and resulting in unwanted foals.
  • Previous attempts to produce eCG in vitro could not maintain activity past 180 days, and peak eCG production came after just 30 days of culture.

Objective of the Study

  • The aim of the study was to establish if organoids derived from the equine chorionic girdle could maintain physiological functionality.
  • Organoids are three-dimensional clusters of cells with potential to self-organize and sustain genetic and phenotypic stability throughout long-term culture.
  • Previous research on human trophoblast organoids indicated successful production and sustained proliferation of human chorionic gonadotropin (hCG), which led the researchers to explore a similar model for eCG production in horses.

Findings from the Study

  • The researchers were successful in generating organoids from the chorionic girdle, marking this as the first instance of such a development.
  • These organoids demonstrated in vitro production of eCG for up to 6 weeks in culture, surpassing previous attempts with explant cultures.
  • The study suggests that equine chorionic girdle organoids could serve as a physiologically representative 3D in vitro model for chorionic girdle development during the early stages of equine pregnancy.
  • This advance presents an alternative method for producing eCG that is less harmful to horse welfare and more sustainable than current practices.

Cite This Article

APA
Thompson RE, Meyers MA, Palmer J, Veeramachaneni DNR, Magee C, de Mestre AM, Antczak DF, Hollinshead FK. (2023). Production of Mare Chorionic Girdle Organoids That Secrete Equine Chorionic Gonadotropin. Int J Mol Sci, 24(11), 9538. https://doi.org/10.3390/ijms24119538

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 24
Issue: 11
PII: 9538

Researcher Affiliations

Thompson, Riley E
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.
Meyers, Mindy A
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.
Palmer, Jennifer
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Veeramachaneni, D N Rao
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Magee, Christianne
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
de Mestre, Amanda M
  • Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Antczak, Douglas F
  • Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Hollinshead, Fiona K
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.

MeSH Terms

  • Pregnancy
  • Humans
  • Horses
  • Animals
  • Female
  • Gonadotropins, Equine / pharmacology
  • Cell Differentiation
  • Trophoblasts
  • Chorionic Gonadotropin / pharmacology
  • Organoids

Grant Funding

  • N/A / Foundation for the Horse

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 43 references
  1. Allen WR, Moor RM. The origin of the equine endometrial cups. I. Production of PMSG by fetal trophoblast cells.. J Reprod Fertil 1972 May;29(2):313-6.
    doi: 10.1530/jrf.0.0290313pubmed: 5023705google scholar: lookup
  2. Wooding FB, Morgan G, Fowden AL, Allen WR. A structural and immunological study of chorionic gonadotrophin production by equine trophoblast girdle and cup cells.. Placenta 2001 Sep-Oct;22(8-9):749-67.
    doi: 10.1053/plac.2001.0707pubmed: 11597196google scholar: lookup
  3. Allen WR. The immunological measurement of pregnant mare serum gonadotrophin.. J Endocrinol 1969 Apr;43(4):593-8.
    doi: 10.1677/joe.0.0430593pubmed: 5767691google scholar: lookup
  4. Allen WR, Hamilton DW, Moor RM. The origin of equine endometrial cups. II. Invasion of the endometrium by trophoblast.. Anat Rec 1973 Dec;177(4):485-501.
    doi: 10.1002/ar.1091770403pubmed: 4762726google scholar: lookup
  5. Wilsher S, Allen WR. Factors influencing equine chorionic gonadotrophin production in the mare.. Equine Vet J 2011 Jul;43(4):430-8.
    doi: 10.1111/j.2042-3306.2010.00309.xpubmed: 21496079google scholar: lookup
  6. Allen WR, Stewart F. Equine placentation.. Reprod Fertil Dev 2001;13(7-8):623-34.
    doi: 10.1071/RD01063pubmed: 11999314google scholar: lookup
  7. Daels PF, Albrecht BA, Mohammed HO. Equine chorionic gonadotropin regulates luteal steroidogenesis in pregnant mares.. Biol Reprod 1998 Nov;59(5):1062-8.
    doi: 10.1095/biolreprod59.5.1062pubmed: 9780310google scholar: lookup
  8. Liu X, Dai Q, Hart EJ, Barrett DM, Rawlings NC, Pierson RA, Bartlewski PM. Ultrasonographic characteristics of ovulatory follicles and associated endocrine changes in cyclic ewes treated with medroxyprogesterone acetate (MAP)-releasing intravaginal sponges and equine chorionic gonadotropin (eCG).. Reprod Domest Anim 2007 Aug;42(4):393-401.
    doi: 10.1111/j.1439-0531.2006.00798.xpubmed: 17635777google scholar: lookup
  9. Lucia T Jr, Corrêa MN, Deschamps JC, Peruzzo IA, Matheus JE, Aleixo JA. Influence of equine chorionic gonadotropin on weaning-to-estrus interval and estrus duration in early-weaned, primiparous, female swine.. J Anim Sci 1999 Dec;77(12):3163-7.
    doi: 10.2527/1999.77123163xpubmed: 10641859google scholar: lookup
  10. De Rensis F, López-Gatius F. Use of equine chorionic gonadotropin to control reproduction of the dairy cow: a review.. Reprod Domest Anim 2014 Apr;49(2):177-82.
    doi: 10.1111/rda.12268pubmed: 24456154google scholar: lookup
  11. Shahneh Z, Sadeghipanah A, Barfourooshi HJ, Emami-Mibody MA. Effects of equine chorionic gonadotropin (eCG) administration and flushing on reproductive performance in Nadooshan goats of Iran.. Afr. J. Biotechnol. 2008;7:3373–3379.
  12. Pierce JG, Parsons TF. Glycoprotein hormones: structure and function.. Annu Rev Biochem 1981;50:465-95.
    doi: 10.1146/annurev.bi.50.070181.002341pubmed: 6267989google scholar: lookup
  13. Sugino H, Bousfield GR, Moore WT Jr, Ward DN. Structural studies on equine glycoprotein hormones. Amino acid sequence of equine chorionic gonadotropin beta-subunit.. J Biol Chem 1987 Jun 25;262(18):8603-9.
    doi: 10.1016/S0021-9258(18)47456-Xpubmed: 3298238google scholar: lookup
  14. Sherman GB, Wolfe MW, Farmerie TA, Clay CM, Threadgill DS, Sharp DC, Nilson JH. A single gene encodes the beta-subunits of equine luteinizing hormone and chorionic gonadotropin.. Mol Endocrinol 1992 Jun;6(6):951-9.
    doi: 10.1210/me.6.6.951pubmed: 1379674google scholar: lookup
  15. Murphy BD. Equine chorionic gonadotrophin: An enigmatic but essential tool.. Anim. Reprod. 2012;9:223–230.
  16. Martinuk SD, Manning AW, Black WD, Murphy BD. Effects of carbohydrates on the pharmacokinetics and biological activity of equine chorionic gonadotropin in vivo.. Biol Reprod 1991 Oct;45(4):598-604.
    doi: 10.1095/biolreprod45.4.598pubmed: 1751634google scholar: lookup
  17. Manteca Vilanova X, De Briyne N, Beaver B, Turner PV. Horse Welfare During Equine Chorionic Gonadotropin (eCG) Production.. Animals (Basel) 2019 Dec 1;9(12).
    doi: 10.3390/ani9121053pmc: PMC6940776pubmed: 31805698google scholar: lookup
  18. Cabrera-Sharp V, Read JE, Richardson S, Kowalski AA, Antczak DF, Cartwright JE, Mukherjee A, de Mestre AM. SMAD1/5 signaling in the early equine placenta regulates trophoblast differentiation and chorionic gonadotropin secretion.. Endocrinology 2014 Aug;155(8):3054-64.
    doi: 10.1210/en.2013-2116pmc: PMC4183921pubmed: 24848867google scholar: lookup
  19. Salman S, Asghar A, Magee C, Winger Q, Bouma G, Bruemmer J. 90 Establishment and characterization of Day 30 equine chorionic girdle and allantochorion cell lines.. Reprod. Fertil. Dev. 2020;32:171.
    doi: 10.1071/RDv32n2Ab90google scholar: lookup
  20. Thway TM, Clay CM, Maher JK, Reed DK, McDowell KJ, Antczak DF, Eckert RL, Nilson JH, Wolfe MW. Immortalization of equine trophoblast cell lines of chorionic girdle cell lineage by simian virus-40 large T antigen.. J Endocrinol 2001 Oct;171(1):45-55.
    doi: 10.1677/joe.0.1710045pubmed: 11572789google scholar: lookup
  21. Gu ZY, Jia SZ, Liu S, Leng JH. Endometrial Organoids: A New Model for the Research of Endometrial-Related Diseases†.. Biol Reprod 2020 Oct 29;103(5):918-926.
    doi: 10.1093/biolre/ioaa124pmc: PMC7609820pubmed: 32697306google scholar: lookup
  22. Lancaster MA, Knoblich JA. Organogenesis in a dish: modeling development and disease using organoid technologies.. Science 2014 Jul 18;345(6194):1247125.
    doi: 10.1126/science.1247125pubmed: 25035496google scholar: lookup
  23. Clevers H. Modeling Development and Disease with Organoids.. Cell 2016 Jun 16;165(7):1586-1597.
    doi: 10.1016/j.cell.2016.05.082pubmed: 27315476google scholar: lookup
  24. Song Y, Fazleabas AT. Endometrial Organoids: A Rising Star for Research on Endometrial Development and Associated Diseases.. Reprod Sci 2021 Jun;28(6):1626-1636.
    doi: 10.1007/s43032-021-00471-zpubmed: 33533008google scholar: lookup
  25. Huch M, Dorrell C, Boj SF, van Es JH, Li VS, van de Wetering M, Sato T, Hamer K, Sasaki N, Finegold MJ, Haft A, Vries RG, Grompe M, Clevers H. In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration.. Nature 2013 Feb 14;494(7436):247-50.
    doi: 10.1038/nature11826pmc: PMC3634804pubmed: 23354049google scholar: lookup
  26. Karthaus WR, Iaquinta PJ, Drost J, Gracanin A, van Boxtel R, Wongvipat J, Dowling CM, Gao D, Begthel H, Sachs N, Vries RGJ, Cuppen E, Chen Y, Sawyers CL, Clevers HC. Identification of multipotent luminal progenitor cells in human prostate organoid cultures.. Cell 2014 Sep 25;159(1):163-175.
    doi: 10.1016/j.cell.2014.08.017pmc: PMC4772677pubmed: 25201529google scholar: lookup
  27. Lancaster MA, Renner M, Martin CA, Wenzel D, Bicknell LS, Hurles ME, Homfray T, Penninger JM, Jackson AP, Knoblich JA. Cerebral organoids model human brain development and microcephaly.. Nature 2013 Sep 19;501(7467):373-9.
    doi: 10.1038/nature12517pmc: PMC3817409pubmed: 23995685google scholar: lookup
  28. Boretto M, Cox B, Noben M, Hendriks N, Fassbender A, Roose H, Amant F, Timmerman D, Tomassetti C, Vanhie A, Meuleman C, Ferrante M, Vankelecom H. Development of organoids from mouse and human endometrium showing endometrial epithelium physiology and long-term expandability.. Development 2017 May 15;144(10):1775-1786.
    doi: 10.1242/dev.148478pubmed: 28442471google scholar: lookup
  29. Turco MY, Gardner L, Hughes J, Cindrova-Davies T, Gomez MJ, Farrell L, Hollinshead M, Marsh SGE, Brosens JJ, Critchley HO, Simons BD, Hemberger M, Koo BK, Moffett A, Burton GJ. Long-term, hormone-responsive organoid cultures of human endometrium in a chemically defined medium.. Nat Cell Biol 2017 May;19(5):568-577.
    doi: 10.1038/ncb3516pmc: PMC5410172pubmed: 28394884google scholar: lookup
  30. Turco MY, Gardner L, Kay RG, Hamilton RS, Prater M, Hollinshead MS, McWhinnie A, Esposito L, Fernando R, Skelton H, Reimann F, Gribble FM, Sharkey A, Marsh SGE, O'Rahilly S, Hemberger M, Burton GJ, Moffett A. Trophoblast organoids as a model for maternal-fetal interactions during human placentation.. Nature 2018 Dec;564(7735):263-267.
    doi: 10.1038/s41586-018-0753-3pmc: PMC7220805pubmed: 30487605google scholar: lookup
  31. Thompson RE, Johnson AK, Dini P, Turco MY, Prado TM, Premanandan C, Burton GJ, Ball BA, Whitlock BK, Pukazhenthi BS. Hormone-responsive organoids from domestic mare and endangered Przewalski's horse endometrium.. Reproduction 2020 Dec;160(6):819-831.
    doi: 10.1530/REP-20-0266pubmed: 33112764google scholar: lookup
  32. Thompson RE, Meyers MA, Pukazhenthi BS, Hollinshead FK. Evaluation of growth, viability, and structural integrity of equine endometrial organoids following cryopreservation.. Cryobiology 2022 Feb;104:56-62.
    doi: 10.1016/j.cryobiol.2021.11.003pubmed: 34788682google scholar: lookup
  33. Thompson RE, Meyers MA, Veeramachaneni DNR, Pukazhenthi BS, Hollinshead FK. Equine Oviductal Organoid Generation and Cryopreservation.. Methods Protoc 2022 Jun 15;5(3).
    doi: 10.3390/mps5030051pmc: PMC9230449pubmed: 35736552google scholar: lookup
  34. de Mestre AM, Bacon SJ, Costa CC, Leadbeater JC, Noronha LE, Stewart F, Antczak DF. Modeling trophoblast differentiation using equine chorionic girdle vesicles.. Placenta 2008 Feb;29(2):158-69.
    doi: 10.1016/j.placenta.2007.10.005pubmed: 18054076google scholar: lookup
  35. Legardinier S, Klett D, Poirier JC, Combarnous Y, Cahoreau C. Mammalian-like nonsialyl complex-type N-glycosylation of equine gonadotropins in Mimic insect cells.. Glycobiology 2005 Aug;15(8):776-90.
    doi: 10.1093/glycob/cwi060pubmed: 15814822google scholar: lookup
  36. Villarraza CJ, Antuña S, Tardivo MB, Rodríguez MC, Mussio P, Cattaneo L, Fontana D, Díaz PU, Ortega HH, Tríbulo A, Macagno A, Bó GA, Ceaglio N, Prieto C. Development of a suitable manufacturing process for production of a bioactive recombinant equine chorionic gonadotropin (reCG) in CHO-K1 cells.. Theriogenology 2021 Sep 15;172:8-19.
    doi: 10.1016/j.theriogenology.2021.05.013pubmed: 34082223google scholar: lookup
  37. Adams AP, Antczak DF. Ectopic transplantation of equine invasive trophoblast.. Biol Reprod 2001 Mar;64(3):753-63.
    doi: 10.1095/biolreprod64.3.753pubmed: 11207188google scholar: lookup
  38. Antczak DF, Oriol JG, Donaldson WL, Poleman C, Stenzler L, Volsen SG, Allen WR. Differentiation molecules of the equine trophoblast.. J Reprod Fertil Suppl 1987;35:371-8.
    pubmed: 3479591
  39. Thompson RE, Johnson AK, Prado TM, Premanandan C, Brown ME, Whitlock BK, Pukazhenthi BS. Dimethyl sulfoxide maintains structure and function of cryopreserved equine endometrial explants.. Cryobiology 2019 Dec;91:90-96.
    doi: 10.1016/j.cryobiol.2019.10.006pubmed: 31626783google scholar: lookup
  40. Thompson RE, Brown ME, Helmick K, Whitlock BK, Pukazhenthi BS. Persian onager (Equus hemionus onager) endometrial explant cryopreservation and in vitro culture.. Anim Reprod Sci 2020 Jun;217:106459.
    doi: 10.1016/j.anireprosci.2020.106459pubmed: 32408971google scholar: lookup
  41. Oriol JG, Poleman JC, Antczak DF, Allen WR. A monoclonal antibody specific for equine trophoblast.. Equine Vet. J. 1989;21:14–18.
    doi: 10.1111/j.2042-3306.1989.tb04665.xgoogle scholar: lookup
  42. Nett TM, Holtan DW, Estergreen VL. Levels of LH, prolactin and oestrogens in the serum of post-partum mares.. J Reprod Fertil Suppl 1975 Oct;(23):201-6.
    pubmed: 1060779
  43. Nett TM, Holtan DW, Estergreen VL. Oestrogens, LH, PMSG, and prolactin in serum of pregnant mares.. J Reprod Fertil Suppl 1975 Oct;(23):457-62.
    pubmed: 1060824

Citations

This article has been cited 6 times.
  1. Mastromonaco G, Mackie P, Russell V, Comizzoli P. Stem Cells and Wildlife Conservation. Adv Exp Med Biol 2026;16:313-339.
    doi: 10.1007/978-3-031-87707-0_10pubmed: 41718875google scholar: lookup
  2. Thompson-Brandhagen RE, Meyers MA, Dunn B, Menjivar NG, Palmer J, Veeramachaneni DNR, Tesfaye D, Hollinshead FK. Characterization of bovine oviductal organoids: Polarity, cryopreservation, hormonal stimulation, and extracellular vesicles. Cell Tissue Res 2026 Jan 16;403(1):5.
    doi: 10.1007/s00441-025-04036-3pubmed: 41543605google scholar: lookup
  3. Thompson RE, Horner AM, Ehresman C, Gad A, Meyers MA, Palmer J, Veeramachaneni DR, Pukazhenthi BS, Hollinshead FK. Canine endometrial organoids respond to exogenous steroid hormones and are an in vitro model for cystic endometrial hyperplasia. Reprod Fertil 2025 Oct 1;6(4).
    doi: 10.1530/RAF-24-0134pubmed: 41187054google scholar: lookup
  4. Pérez MM, Veronez LC, Cordeiro FA, de Castro Figueiredo Bordon K, Arantes EC, Munari C, Marquele-Oliveira F, Muller V, Guimarães DA. An Alternative In Vitro Methodology for the Quality Control of Equine-Chorionic Gonadotropin in Commercial Products. ACS Omega 2025 Feb 25;10(7):7211-7219.
    doi: 10.1021/acsomega.4c10780pubmed: 40028073google scholar: lookup
  5. Byambaragchaa M, Park SH, Park MH, Kang MH, Min KS. Enhanced Production and Functional Characterization of Recombinant Equine Chorionic Gonadotropin (rec-eCG) in CHO-DG44 Cells. Biomolecules 2025 Feb 14;15(2).
    doi: 10.3390/biom15020289pubmed: 40001592google scholar: lookup
  6. Mu S, Shen Y, Ren H, Ulaangerel T, Yi M, Zhao B, Hao A, Liu Q, Wen X, Dugarjaviin M, Bou G. Effects of Interleukin-6 (IL-6) on In Vitro Cultured Equine Chorionic Girdle Cells. Animals (Basel) 2025 Feb 6;15(3).
    doi: 10.3390/ani15030450pubmed: 39943220google scholar: lookup
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