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Addition of zinc to human ejaculate prior to cryopreservation prevents freeze-thaw-induced DNA damage and preserves sperm function

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Abstract

Purpose

To study the effect of addition of zinc to human semen sample prior to cryopreservation on post-thaw sperm quality and function.

Methods

Semen samples were collected from men attending university infertility clinic for semen analysis (n=109). Liquefied semen samples were cryopreserved in glycerol-egg yolk- citrate medium with or without the prior addition of zinc (100 μM) and stored in liquid nitrogen. After 10 days, the semen samples were thawed to assess the outcome. Sperm motility, DNA integrity, mitochondrial potential and the ability of spermatozoa to undergo capacitation and acrosome reaction was assessed in post-thaw samples.

Results

Semen samples cryopreserved after addition of zinc had a significantly higher percentage of sperm with intact DNA (p<0.001), mitochondrial function (p<0.001) and progressive motility (p<0.01) compared to the semen samples cryopreserved without zinc supplementation. Apart from this, ability to undergo capacitation and acrosome reaction in vitro was significantly higher in semen samples cryopreserved with zinc (p<0.0001 and p<0.001 respectively).

Conclusions

Addition of zinc to semen samples prior to cryopreservation helps in preventing the freeze-thaw-induced sperm DNA damage and loss of sperm function.

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Acknowledgments

Authors acknowledge financial assistance from Kasturba Medical College, Manipal, India, in the form of Postgraduate thesis grant sanctioned to APK (Grant No. PGR042). Technical support from Ms. Jayalaxmi Pai and Ms. Kirthi Patil (Clinical Embryology, Kasturba Medical College, Manipal) and Ms. Gimna Rathesh (Division of Nutrition, St. John’s Research Institute, St. John’s National Academy of Health Sciences, Bangalore) is thankfully acknowledged. Authors thank Dr. Prashanth Naik, Mangalore University for critical editing of this manuscript.

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Authors and Affiliations

  1. Clinical Embryology, Department of Obstetrics and Gynecology, Kasturba Medical College, Manipal University, Manipal, 576 104, India

    Aditi P. Kotdawala, Sangeetha Kumar, Sujith R. Salian, Pratap Kumar, Guruprasad Kalthur & Satish K. Adiga

  2. Division of Nutrition, St. John’s Research Institute, St. John’s National Academy of Health Sciences, Bangalore, India

    Prashanth Thankachan & Kaushik Govindraj

Authors
  1. Aditi P. Kotdawala
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  2. Sangeetha Kumar
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  3. Sujith R. Salian
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  4. Prashanth Thankachan
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  5. Kaushik Govindraj
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  6. Pratap Kumar
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  7. Guruprasad Kalthur
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  8. Satish K. Adiga
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Corresponding author

Correspondence to Guruprasad Kalthur.

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Capsule

Addition of zinc to liquefied ejaculate prior to cryopreservation using glycerol- egg yolk- citrate buffered medium protects spermatozoa from freeze-thaw-induced DNA damage and loss of sperm function.

Author contribution

APK, SK, KG and SRS performed the study (APK and SK had equal contribution)

GK designed the study and wrote the paper

PT, SKA and PK helped in analyzing the data and writing the paper

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Kotdawala, A.P., Kumar, S., Salian, S.R. et al. Addition of zinc to human ejaculate prior to cryopreservation prevents freeze-thaw-induced DNA damage and preserves sperm function. J Assist Reprod Genet 29, 1447–1453 (2012). https://doi.org/10.1007/s10815-012-9894-8

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  • DOI: https://doi.org/10.1007/s10815-012-9894-8

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