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Home / Equine Research Bank / Cryobiology / Evaluation of growth, viability, and structural integrity of...
Cryobiology2021; 104; 56-62; doi: 10.1016/j.cryobiol.2021.11.003

Evaluation of growth, viability, and structural integrity of equine endometrial organoids following cryopreservation.

Abstract: Reproductive diseases in mares are a significant cause of subfertility and profound economic loss in the equine industry. Utilizing a 3D in vitro cell culture system that recapitulates the in vivo physiology will reduce time, cost, and welfare concerns associated with in vivo reproductive research in mares. If this 3D model is combined with effective cryopreservation, reproductive research on mares can occur year-round, which is not currently possible in this seasonal species. Endometrial organoids, 3D in vitro cell clusters that exhibit in vivo uterine physiology, have been established in mice, women, and mares. Here we report the first comprehensive assessment of cryopreservation of endometrial organoids in the domestic mare. Organoid growth rate was not affected by the type of freezing media. However, growth rate varied among non-cryopreserved controls, organoids cryopreserved at passage 0 (P0), and organoids cryopreserved at passage 3 (P3). Additionally, there was no difference in organoid viability among freezing media or freezing timepoint (passages). Furthermore, fresh and frozen-thawed organoids displayed positive immunohistochemical staining for ZO-1, which is a marker for intercellular tight junctions, and for periodic acid-Schiff staining as marker for organoid function through mucin production. Results demonstrate that equine endometrial organoids can be cryopreserved with 10% dimethyl sulfoxide with minimal detrimental effects while maintaining intercellular tight junctions (ZO-1) and secretory function. Availability of cryopreserved endometrial organoids may permit expanded research on uterine pathologies that negatively affect mare fertility and improve efficiency, reduce cost, and minimize animal welfare concerns associated with in vivo research in the domestic mare.
Copyright © 2021 Elsevier Inc. All rights reserved.
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Publication Date: 2021-11-14 PubMed ID: 34788682DOI: 10.1016/j.cryobiol.2021.11.003Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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.

The research paper explores the effects of cryopreservation on equine endometrial organoids, 3D cell clusters that mimic in vivo uterine physiology. It reveals that the organoids can be effectively frozen for later use without damaging their functional and structural properties.

Study Purpose and Methodology

  • The research aims to assess the feasibility of cryopreserving equine endometrial organoids with the purpose of facilitating year-round reproduction studies on mares.
  • Cryopreservation, defined as the freezing of biological material for future use, could eliminate the limitations of conducting research within a specific reproductive season.
  • Endometrial organoids are used as they mimic the in vivo uterine physiology more accurately than traditional 2D cell culture methods.
  • The study measures the growth rate, viability and structural integrity of these organoids after cryopreservation and thawing.

Key Findings

  • Variations in growth rate were observed among non-cryopreserved controls, and organoids cryopreserved at two different stages, known as passage 0 (P0) and passage 3 (P3).
  • Regardless of the type of freezing medium used, the growth rate was not affected.
  • There was no significant difference in organoid viability among different freezing media or freezing stages.
  • Both fresh and frozen-thawed organoids positively stained for ZO-1, an indication of intercellular tight junctions, and periodic acid-Schiff, a marker of mucin production and organoid function.
  • The experiment successfully verified that equine endometrial organoids could be cryopreserved with 10% dimethyl sulfoxide, a common cryoprotectant, with minimal adverse effects.

Implications of Results

  • The ability to maintain healthy and functional equine endometrial organoids post-cryopreservation is a significant research breakthrough.
  • The results could enable expanded study on uterine pathologies, which could subsequently lead to improved fertility treatments for mares.
  • The ability to conduct reproductive research out of season, made possible by cryopreservation, will reduce costs, increase efficiency, and address animal welfare issues often associated with in vivo research.

Cite This Article

APA
Thompson RE, Meyers MA, Pukazhenthi BS, Hollinshead FK. (2021). Evaluation of growth, viability, and structural integrity of equine endometrial organoids following cryopreservation. Cryobiology, 104, 56-62. https://doi.org/10.1016/j.cryobiol.2021.11.003

Publication

Cryobiology
ISSN: 1090-2392
NlmUniqueID: 0006252
Country: Netherlands
Language: English
Volume: 104
Pages: 56-62
PII: S0011-2240(21)00178-4

Researcher Affiliations

Thompson, Riley E
  • Colorado State University, Department of Clinical Sciences, 1601 Campus Delivery, Fort Collins, CO, 80523, USA. Electronic address: riley.thompson2@colostate.edu.
Meyers, Melinda A
  • Colorado State University, Department of Clinical Sciences, 1601 Campus Delivery, Fort Collins, CO, 80523, USA.
Pukazhenthi, Budhan S
  • Smithsonian Conservation Biology Institute, Center for Species Survival, 1500 Remount Road, Front Royal, VA, 22630, USA.
Hollinshead, Fiona K
  • Colorado State University, Department of Clinical Sciences, 1601 Campus Delivery, Fort Collins, CO, 80523, USA.

MeSH Terms

  • Animals
  • Cryopreservation / methods
  • Dimethyl Sulfoxide / metabolism
  • Dimethyl Sulfoxide / pharmacology
  • Endometrium / physiology
  • Female
  • Horses
  • Humans
  • Mice
  • Organoids / metabolism
  • Uterus

Citations

This article has been cited 12 times.
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  8. Zolfaghar M, Acharya P, Joshi P, Choi NY, Shrestha S, Lekkala VKR, Kang SY, Lee M, Lee MY. Cryopreservation of neuroectoderm on a pillar plate and in situ differentiation into human brain organoids. bioRxiv 2024 Jul 25;.
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  10. Gabriel V, Zdyrski C, Sahoo DK, Ralston A, Wickham H, Bourgois-Mochel A, Ahmed B, Merodio MM, Paukner K, Piñeyro P, Kopper J, Rowe EW, Smith JD, Meyerholz D, Kol A, Viall A, Elbadawy M, Mochel JP, Allenspach K. Adult Animal Stem Cell-Derived Organoids in Biomedical Research and the One Health Paradigm. Int J Mol Sci 2024 Jan 5;25(2).
    doi: 10.3390/ijms25020701pubmed: 38255775google scholar: lookup
  11. Menjivar NG, Gad A, Thompson RE, Meyers MA, Hollinshead FK, Tesfaye D. Bovine oviductal organoids: a multi-omics approach to capture the cellular and extracellular molecular response of the oviduct to heat stress. BMC Genomics 2023 Oct 27;24(1):646.
    doi: 10.1186/s12864-023-09746-ypubmed: 37891479google scholar: lookup
  12. Thompson RE, Meyers MA, Palmer J, Veeramachaneni DNR, Magee C, de Mestre AM, Antczak DF, Hollinshead FK. Production of Mare Chorionic Girdle Organoids That Secrete Equine Chorionic Gonadotropin. Int J Mol Sci 2023 May 31;24(11).
    doi: 10.3390/ijms24119538pubmed: 37298490google scholar: lookup
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