Human sperm DNA integrity : A difference of storage temperature and period of freeze-drying process

Authors

  • silvia werdhy lestari Universitas Indonesia
  • ahmad silalahi Andrology Specialist Program, Medical Faculty of the University of Airlangga / Dr. Soetomo Hospital Surabaya, Indonesia
  • hamdani lunardi Medical Biology Department, Medical Faculty of the University of Airlangga, Surabaya, Indonesia.
  • agustinus agustinus Medical Biology Department, Medical Faculty of the University of Airlangga, Surabaya, Indonesia.

Abstract

The cryopreserve technique using liquid nitrogen is used to preserve human sperm. Nevertheless, this technique has several disadvantages, such as bacterial/viral contamination and its expensive cost. Freeze-drying process offers more simple and cheaper of sperm storage technique. This study aimed to investigate the effect of storage temperature and period of freeze-dried human sperm on DNA integrity. Human sperm samples of 15 donors were prepared with simple washing techniques. Post-washing samples (T0) were divided into 4 groups with the same volume. The four group of samples carried on the freeze-drying process and stored based on different temperature and period: 4°C for 1 week (T1), room temperature (24-25ºC) for 1 week (T2), 4°C for 3 months (T3), and room temperature for 3 months (T4). Sperm DNA integrity was analyzed before and after the storage period of each group. There was a statistically significant difference of sperm DNA integrity of freezes - dried at 4°C in 1 week versus 3 months (T1 vs T3), at room temperature in 1 week versus 3 months (T2 vs T4) (p <0.05) and between both temperatures in 3 months (T3 vs T4) (p<0.05). In conclusion, temperature at 4°C and period in 1 week is the best temperature and storage to store freeze-dried human sperm, based on sperm DNA integrity. Keywords: freeze-drying process, human sperm, sperm DNA integrity

Author Biographies

silvia werdhy lestari, Universitas Indonesia

Departemen Biologi Kedokteran Fakultas Kedokteran Universitas Indonesia

ahmad silalahi, Andrology Specialist Program, Medical Faculty of the University of Airlangga / Dr. Soetomo Hospital Surabaya, Indonesia

Andrology Specialist Program, Medical Faculty of the University of Airlangga / Dr. Soetomo Hospital Surabaya, Indonesia

hamdani lunardi, Medical Biology Department, Medical Faculty of the University of Airlangga, Surabaya, Indonesia.

Medical Biology Department, Medical Faculty of the University of Airlangga, Surabaya, Indonesia.

agustinus agustinus, Medical Biology Department, Medical Faculty of the University of Airlangga, Surabaya, Indonesia.

Medical Biology Department, Medical Faculty of the University of Airlangga, Surabaya, Indonesia.

References

Raja DR, Natrajamani S. SPERM DNA INTEGRITY BEFORE AND AFTER CRYOPRESERVATION. 2013;2(4):15.

Le MT, Nguyen TTT, Nguyen TT, Nguyen TV, Nguyen TAT, Nguyen QHV, et al. Does conventional freezing affect sperm DNA fragmentation? Clin Exp Reprod Med. 2019 Jun 1;46(2):67–75.

Cankut S, Dinc T, Cincik M, Ozturk G, Selam B. Evaluation of Sperm DNA Fragmentation via Halosperm Technique and TUNEL Assay Before and After Cryopreservation. Reprod Sci. 2019 Dec 1;26(12):1575–81.

Joaquim DC, Borges ED, Viana IGR, Navarro PA, Vireque AA. Risk of Contamination of Gametes and Embryos during Cryopreservation and Measures to Prevent Cross-Contamination [Internet]. BioMed Research International. 2017 [cited 2019 Mar 20]. Available from: https://www.hindawi.com/journals/bmri/2017/1840417/

Molina I, Mari M, Martínez JV, Novella-Maestre E, Pellicer N, Pemán J. Bacterial and fungal contamination risks in human oocyte and embryo cryopreservation: open versus closed vitrification systems. Fertility and Sterility. 2016 Jul;106(1):127–32.

Nangia AK, Krieg SA, Kim SS. Clinical guidelines for sperm cryopreservation in cancer patients. Fertility and Sterility. 2013 Nov 1;100(5):1203–9.

Rozati H, Handley T, Jayasena C. Process and Pitfalls of Sperm Cryopreservation. J Clin Med [Internet]. 2017 Sep 19 [cited 2019 Mar 25];6(9). Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615282/

Keskintepe L, Eroglu A. Freeze-drying of mammalian sperm. Methods in molecular biology (Clifton, NJ). 2015 Jan 1;1257:489–97.

Patrizio P, Natan Y, Barak Y, Levi Setti P, Arav A. A simple new method for the freeze-drying and storage of human sperm. Fertility and Sterility. 2016 Sep;106(3):e307.

Gianaroli L, Magli MC, Stanghellini I, Crippa A, Crivello AM, Pescatori ES, et al. DNA integrity is maintained after freeze-drying of human spermatozoa. Fertility and Sterility. 2012 May;97(5):1067-1073.e1.

Pramesti MB, Lunardhi H, I’tishom R. HUMAN SPERM PRESERVATION USING FREEZE-DRYING An EXPERIMENTAL LABORATORY STUDY. 2010;9.

World Health Organization, editor. WHO laboratory manual for the examination and processing of human semen. 5th ed. Geneva: World Health Organization; 2010. 271 p.

Kusakabe H. Chromosomal integrity and DNA damage in freeze-dried spermatozoa. Reprod Med Biol. 2011 Dec;10(4):199–210.

Olaciregui M, Gil L. Freeze-dried spermatozoa: A future tool? Reproduction in Domestic Animals. 2017 Apr;52:248–54.

Peitao W, Cuihua S, Daqing S, Li Q, Liu Z, Xiuyan F. The Effect of Freeze Drying on the Deoxyribonucleic Acid Chain of Human Sperm. In: 2008 2nd International Conference on Bioinformatics and Biomedical Engineering [Internet]. Shanghai, China: IEEE; 2008 [cited 2019 Oct 14]. p. 919–23. Available from: http://ieeexplore.ieee.org/document/4535106/

Domingo P, Olaciregui M, González N, De Blas I, Gil L. Long-term preservation of freeze-dried rabbit sperm by adding rosmarinic acid and different chelating agents. Cryobiology. 2018;81:174–7.

Zhang M, Oldenhof H, Sydykov B, Bigalk J, Sieme H, Wolkers WF. Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity. Scientific Reports. 2017 Jul 24;7(1):6198.

Published

2021-02-20
Statistics
Abstract Display: 377
PDF Downloads: 270

Issue

Volume 12 Issue 09 (2020) Sept. 2020

Section

Articles