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Detailed cell-level analysis of sperm nuclear quality among the different hypo-osmotic swelling test (HOST) classes

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

We studied the quality differences between the different hypo-osmotic swelling test (HOST) classes, as measured by criteria of DNA fragmentation, DNA decondensation, and nuclear architecture. The aim was to find particular HOST classes associated with good-quality metrics, which may be potentially used in ICSI (intra-cytoplasmic sperm injection).

Methods

Ten patients from the Department of Reproductive Medicine at Tenon Hospital (Paris, France) were included. Their semen samples were collected and divided into two fractions: one was incubated in a hypo-osmotic solution as per HOST protocol and sorted by sperm morphology, and a second was incubated without undergoing the HOST protocol to serve as an unsorted baseline. Three parameters were assessed: DNA fragmentation (TUNEL assay), DNA decondensation (chromomycin A3 assay), and nuclear architecture (FISH, with telomeric and whole chromosome painting probes). The different HOST classes were evaluated for these three parameters, and statistical analysis was performed for each class versus the unsorted non-HOST-treated sperm. Results with p<0.05 were considered statistically significant.

Results

For each of the parameters evaluated, we found significant differences between HOST-selected spermatozoa and non-selected spermatozoa. Overall, spermatozoa of HOST classes B and B+ exhibited the highest quality based on four metrics (low DNA fragmentation, low DNA decondensation, short inter-telomeric distance, and small chromosome 1 territory area), while spermatozoa of HOST classes A and G exhibited the poorest quality by these metrics.

Conclusion

In addition to their pathophysiological interest, our results open possibilities of sperm selection prior to ICSI, which may allow for optimization of reproductive outcomes in heretofore unstudied patient populations.

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Funding

No specific funding was sought for this study, and departmental funds were used to support the authors (Sorbonne Université, INSERM, Maladies génétiques d’expression pédiatrique, APHP, Hôpital d’Enfants Armand Trousseau, Département de Génétique Médicale, F-75012 Paris, France).

Author information

Author notes

  1. Adrien Bloch, Cynthia Nicolas, and Eli Rogers equally contributed to this work as first authors.

Authors and Affiliations

  1. Département de Génétique Médicale, Sorbonne Université, INSERM, Maladies génétiques d’expression pédiatrique, APHP, Hôpital d’Enfants Armand Trousseau, 26 avenue du Dr Arnold Netter, F-75012, Paris, France

    Adrien Bloch, Eli J. Rogers, Cynthia Nicolas, Hélène Morel, Jean-Pierre Siffroi & Alexandre Rouen

  2. Centre de Génétique Médicale Alpigène, Synlab France, Lyon, France

    Tanguy Martin-Denavit

  3. mojo (Nanovare SAS), Lyon, France

    Miguel Monteiro & Daniel Thomas

  4. Service de Biologie de la Reproduction CECOS, Hôpital Tenon (AP-HP), Sorbonne-Université, 75020, Paris, France

    Rachel Lévy & Charlotte Dupont

  5. Centre de recherche Saint-Antoine, Inserm US938, Sorbonne Université, 75012, Paris, France

    Rachel Lévy & Charlotte Dupont

Authors
  1. Adrien Bloch
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  2. Eli J. Rogers
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  3. Cynthia Nicolas
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  4. Tanguy Martin-Denavit
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  5. Miguel Monteiro
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  6. Daniel Thomas
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  7. Hélène Morel
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  8. Rachel Lévy
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  9. Jean-Pierre Siffroi
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  10. Charlotte Dupont
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  11. Alexandre Rouen
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Contributions

Adrien Bloch, Eli Rogers, and Cynthia Nicolas performed microscope analysis, and participated in recruiting subjects and in writing the manuscript.

Tanguy Martin-Denavit performed the chromomycin A3 technique.

Miguel Monteiro and Daniel Thomas provided expertise on the figures and on sperm selection possibilities.

Hélène Morel participated in microscope analysis.

Rachel Levy and Jean-Pierre Siffroi participated in conceiving the study.

Charlotte Dupont participated in recruiting the subjects and performed the initial routine sperm analysis.

Alexandre Rouen conceived and coordinated the study.

Corresponding author

Correspondence to Alexandre Rouen.

Ethics declarations

Ethics approval

The ethics commission CECOS (Centre d’Etude et de Conservation des Oeufs et du Sperm humains) considered this project to be exempted from IRB approval since it did not involve any additional medical intervention and solely used remnants from sperm samples obtained as part of the routine clinical care

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Informed consent was obtained from each subject.

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All authors haven consented for publication of this manuscript.

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The authors declare no competing interests.

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Bloch, A., Rogers, E.J., Nicolas, C. et al. Detailed cell-level analysis of sperm nuclear quality among the different hypo-osmotic swelling test (HOST) classes. J Assist Reprod Genet 38, 2491–2499 (2021). https://doi.org/10.1007/s10815-021-02232-y

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  • DOI: https://doi.org/10.1007/s10815-021-02232-y

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