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The effect of cryopreservation technique in a very low number sperm count at Soetomo Hospital, Surabaya, Indonesia

  • Rossy Sintya Marthasari ,
  • Agustinus Agustinus ,
  • Supardi Supardi ,
  • Aucky Hinting ,
  • Reny I’tishom ,


Background: The massive development of Assisted Reproductive Technology (ART) offers new hope for the infertile couple. However, the advanced storage methods to preserve the quality of sperm has been raising by cryopreservation technique. This study aims to analyze the motility rate, recovery rate, and viability in subgroups of cryopreserved spermatozoa from the oligozoospermic patient as the quality parameter.

Method: A true-experimental study by pre-test and post-test group design was conducted among 13 oligozoospermic patients with a very low sperm counts at Medical Biology Laboratory Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia. Parameters assessed in this study include motility, viability, and concentration of spermatozoa according to Laboratory Manual for the Examination and Processing of Human Sperm 5th edition. The samples were divided into 4 groups such as Group 1 (71-100 sperm), Group 2 (31-70 sperm), Group 3 (11-30 sperm), and Group 4 (1-10 sperm) based on sperm counts which underwent 30 repetitions of cryopreservation. Data were analyzed using SPSS version 17 for Windows. 

Results: There was a significant difference in the recovery rate among Group 1 (34.88±1.87), Group 2 (44.77±1.89), Group 3 (61.83±2.09), and Group 4 (70.90±2.71) of cryopreservation technique (P<0.001). However, there was no significantly different in the motility rate of sperm-cryopreservation technique among Group 1 (65.60±14.63), Group 2 (52.50±26.28), Group 3 (39.90±9.95), and Group 4 (44.70±41.77) (p=0.070). Based on the sperm-viability parameter assessed in this study, there was no significant difference among Group 1 (56.40±3.89), Group 2 (58.60±5.99), Group 3 (59.60±9.96), and Group 4 (56.70±11.30) (p=0.504).

Conclusions: The sperm-cryopreservation technique significantly affected the recovery rate of very low sperm number count among groups. However, there was no significantly different in the motility rate and viability parameter in this study. 


  1. AbdelHafez F, Bedaiwy M, El-Nashar SA, Sabanegh E, Desai N. Techniques for cryopreservation of individual or small numbers of human spermatozoa: a systematic review. Hum Reprod Update. 2009;15(2):153â€164.
  2. Huang JY, Rosenwaks Z. Assisted reproductive techniques. Methods Mol Biol. 2014;1154:171â€231.
  3. Le MT, Nguyen TTT, Nguyen TT, et al. Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects. Eur J Obstet Gynecol Reprod Biol. 2019;234:14â€20.
  4. Koscinski I, Wittemer C, Lefebvre-Khalil V, Marcelli F, Defossez A, Rigot JM. Optimal management of extreme oligozoospermia by an appropriate cryopreservation programme. Hum Reprod. 2007;22(10):2679â€2684.
  5. Ashrafi M, Jahanian Sadatmahalleh S, Akhoond MR, Ghaffari F, Zolfaghari Z. ICSI Outcome in Infertile Couples with Different Causes of Infertility: A Cross-Sectional Study. Int J Fertil Steril. 2013;7(2):88â€95.
  6. Kathrins M, Abhyankar N, Shoshany O, et al. Post-thaw recovery of rare or very low concentrations of cryopreserved human sperm. Fertil Steril. 2017;107(6):1300â€1304.
  7. Moshe W, Prins GS. Sperm banking: Indication and techniques. In: Lipshultz LI, Howards SS, Niederberger CS. Infertility in the Male. 4th edition. Cambridge University Press New York. 2009:593-602
  8. Henkel RR, Schill WB. Sperm preparation for ART. Reprod Biol Endocrinol. 2003;1:108.
  9. Hinting A, Lunardhi H. Better sperm selection for intracytoplasmic sperm injection with the side migration technique. Andrologia. 2001;33(6):343â€346.
  10. Vutyavanich T, Lattiwongsakorn W, Piromlertamorn W, Samchimchom S. Repeated vitrification/warming of human sperm gives better results than repeated slow programmable freezing. Asian J Androl. 2012;14(6):850â€854.
  11. Tavukcuoglu S, Al-Azawi T, Khaki AA, Al-Hasani S. Is vitrification standard method of cryopreservation. Middle East Fertility Society Journal. 2012;17(3)152-156
  12. Agha-Rahimi A, Khalili MA, Nabi A, Ashourzadeh S. Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding. Reprod Biomed Online. 2014;28(3):352â€358.
  13. Riva NS, Ruhlmann C, Iaizzo RS, Marcial López CA, Martínez AG. Comparative analysis between slow freezing and ultra-rapid freezing for human sperm cryopreservation. JBRA Assist Reprod. 2018;22(4):331â€337.
  14. Schuster TG, Keller LM, Dunn RL, Ohl DA, Smith GD. Ultra-rapid freezing of very low numbers of sperm using cryoloops. Hum Reprod. 2003;18(4):788â€795.
  15. Hossain AM, Osuamkpe CO. Sole use of sucrose in human sperm cryopreservation. Arch Androl. 2007;53(2):99â€103.
  16. Isachenko E, Isachenko V, Weiss JM, Kreienberk R, Katkov II, Schulz M, et al. Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose. Reproduction. 2008;136(2):167â€173.
  17. Rozati H, Handley T, Jayasena CN. Process and Pitfalls of Sperm Cryopreservation. J Clin Med. 2017;6(9):89.
  18. Nallella KP, Sharma RK, Allamaneni SS, Aziz N, Agarwal A. Cryopreservation of human spermatozoa: comparison of two cryopreservation methods and three cryoprotectants. Fertil Steril. 2004;82(4):913â€918.

How to Cite

Marthasari, R. S., Agustinus, A., Supardi, S., Hinting, A., & I’tishom, R. (2020). The effect of cryopreservation technique in a very low number sperm count at Soetomo Hospital, Surabaya, Indonesia. Bali Medical Journal, 9(1), 371–375.




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Rossy Sintya Marthasari
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Agustinus Agustinus
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Supardi Supardi
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Aucky Hinting
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Reny I’tishom
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