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Andrology2017; 5(2); 336-346; doi: 10.1111/andr.12308

Xenografting of isolated equine (Equus caballus) testis cells results in de novo morphogenesis of seminiferous tubules but not spermatogenesis.

Abstract: The study of spermatogenesis in the horse is challenging because of the absence of an in vitro system that is capable of reproducing efficient spermatogenesis and because of the difficulties and costs associated with performing well-controlled studies in vivo. In an attempt to develop novel methods for the study of equine spermatogenesis, we tested whether cells from enzymatically digested pre-pubertal equine testicular tissue were capable of de novo tissue formation and spermatogenesis following xenografting under the back skin of immunocompromised mice. Testes were obtained from normal pre-pubertal colts and dissociated into cell suspensions using trypsin/collagenase digestion. Resulting cell pellets, consisting of both somatic and germ cells, were injected into fascial pockets under the back skin of immunocompromised, castrated mice and maintained for between 1 and 14 months. Mice were killed and grafts were recovered and analyzed. As has been reported for testis cell suspensions from pigs, mice, cattle, and sheep, de novo formation of equine testicular tissue was observed, as evidenced by the presence of seminiferous tubules and an interstitial compartment. There was an increased likelihood of de novo testicular formation as grafting period increased. Using indirect immunofluorescence, we confirmed the presence of spermatogonia in de novo formed seminiferous tubules. However, we found no evidence of meiotic or haploid cells. These results indicate that dissociated pre-pubertal equine testis cells are capable of reorganizing into the highly specialized endocrine and spermatogenic compartments of the testis following ectopic xenografting. However, in spite of the presence of spermatogonia within the seminiferous tubules, spermatogenesis does not occur. Although this technique does allow access to the cells within the seminiferous tubule and interstitial compartments of the equine testis prior to reaggregation, the absence of spermatogenesis will limit its use as a method for the study of testicular function in the horse.
© 2017 American Society of Andrology and European Academy of Andrology.
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Publication Date: 2017-02-03 PubMed ID: 28160442PubMed Central: PMC6018012DOI: 10.1111/andr.12308Google 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 looks into a novel method to observe spermatogenesis in horses, involving xenografting cells from pre-pubertal equine testes into immunocompromised mice. The cells were observed to form new testicular tissue structure, but did not undergo spermatogenesis.

Study Design and Methods

  • The study challenges were based on the absence of an in vitro system that can reproduce efficient spermatogenesis and the difficulties associated with performing well-controlled in vivo studies due to cost. The objective was to find a new way to study equine spermatogenesis.
  • The researchers used pre-pubertal equine testes, which were dissociated into cell suspensions via digestion with trypsin/collagenase. The cell suspensions, made up of both somatic and germ cells, were then injected into immunocompromised, castrated mice. The mice were maintained for a duration between 1 and 14 months.
  • After which, the mice were killed, and the grafts were recovered for analysis. The hypothesis was that these cells would undergo de novo tissue formation and spermatogenesis, meaning they would reform into new tissue and begin producing sperm cells once grafted into the mice.

Results

  • The results showed that de novo formation of equine testicular tissue did occur, which is similar to what has been observed in pigs, mice, cattle, and sheep. This was indicated by the presence of seminiferous tubules and an interstitial compartment within the grafts.
  • Further, the results indicated a higher chance of de novo testicular formation as the grafting period increased, providing evidence of the potential for these cells to reform into the highly specialized endocrine and spermatogenic compartments of the testis.
  • Despite these discoveries, no signs of meiotic or haploid cells were found, meaning no evidence for the occurrence of actual spermatogenesis was observed.

Conclusion

  • Although dissociated pre-pubertal equine testes cells were capable of reorganizing into the key compartments of testis following xenografting, no spermatogenesis occurred. As a result, while this technique presents an opportunity to access the cells within these testicular compartments prior to their reaggregation, its potential use in studying equine testicular function is limited due to the absence of spermatogenesis.

Cite This Article

APA
Zeng W, Alpaugh W, Stefanovski D, Schlingmann K, Dobrinski I, Turner RM. (2017). Xenografting of isolated equine (Equus caballus) testis cells results in de novo morphogenesis of seminiferous tubules but not spermatogenesis. Andrology, 5(2), 336-346. https://doi.org/10.1111/andr.12308

Publication

Andrology
ISSN: 2047-2927
NlmUniqueID: 101585129
Country: England
Language: English
Volume: 5
Issue: 2
Pages: 336-346

Researcher Affiliations

Zeng, W
  • College of Animal Science and Technology, Northwest A&F University, Yangling, China.
Alpaugh, W
  • Department of Comparative Biology & Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.
Stefanovski, D
  • Department of Clinical Studies, New Bolton Center, University of Pennsylvania School of Veterinary Medicine, Kennett Square, PA, USA.
Schlingmann, K
  • Department of Clinical Studies, New Bolton Center, University of Pennsylvania School of Veterinary Medicine, Kennett Square, PA, USA.
Dobrinski, I
  • Department of Comparative Biology & Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.
Turner, R M
  • Department of Clinical Studies, New Bolton Center, University of Pennsylvania School of Veterinary Medicine, Kennett Square, PA, USA.

MeSH Terms

  • Animals
  • Horses
  • Male
  • Mice
  • Morphogenesis / physiology
  • Seminiferous Tubules / growth & development
  • Spermatogenesis / physiology
  • Spermatogonia / transplantation
  • Testis / cytology
  • Transplantation, Heterologous

Grant Funding

  • R01 OD016575 / NIH HHS

Conflict of Interest Statement

. The authors declare no conflicts of interest relevant to this publication.

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Citations

This article has been cited 1 times.
  1. Lima DBC, Silva LDMD, Comizzoli P. Influence of warming and reanimation conditions on seminiferous tubule morphology, mitochondrial activity, and cell composition of vitrified testicular tissues in the domestic cat model. PLoS One 2018;13(11):e0207317.
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