Volume 22, Issue 4 (April 2024)                   IJRM 2024, 22(4): 305-316 | Back to browse issues page

Ethics code: H2019/010

XML Persian Abstract Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Le M T, Van Nguyen T, Nguyen T T T, Dang H N T, Nguyen Q H V. Impact of cryoprotectant-free sperm vitrification in pulled-glass capillary on sperm parameters and DNA integrity: A lab trial study. IJRM 2024; 22 (4) :305-316
URL: http://ijrm.ir/article-1-3061-en.html
Impact of cryoprotectant-free sperm vitrification in pulled-glass capillary on sperm parameters and DNA integrity: A lab trial study
Minh Tam Le * 1, Trung Van Nguyen2 , Thai Thanh Thi Nguyen2 , Hong Nhan Thi Dang2 , Quoc Huy Vu Nguyen3
1- Center for Reproductive Endocrinology and Infertility, Hue University of Medicine and Pharmacy, Hue University, Hue City, Vietnam. Department of Obstetrics and Gynecology, Hue University of Medicine and Pharmacy, Hue University, Hue City, Vietnam. , leminhtam@huemed-univ.edu.vn
2- Center for Reproductive Endocrinology and Infertility, Hue University of Medicine and Pharmacy, Hue University, Hue City, Vietnam.
3- Department of Obstetrics and Gynecology, Hue University of Medicine and Pharmacy, Hue University, Hue City, Vietnam.
Abstract:   (365 Views)
Background: Vitrification is a recently introduced yet widely applied assisted reproduction technique. So far, the effects of the chemicals and devices in vitrification on sperm motility and DNA integrity are still unclear.
Objective: This study aimed to examine sperm quality, as determined by semen analysis and sperm DNA integrity when vitrified with or without cryoprotectant agents (CPAs) using pulled-glass capillaries.
Materials and Methods: Between February and June 2020, 50 infertile men from the Hue Center for Reproductive Endocrinology and Infertility, Hue University of Medicine and Pharmacy, Vietnam, were enrolled. Sperm samples, prepared using the swim-up technique, were divided into 2 groups: vitrification with CPAs (group 1) and without CPAs (group 2). Vitrified sperm samples were preserved in 10 µL pulled-glass capillaries. Motility, sperm membrane integrity, and the DNA fragmentation index were tested.
Results: Sperm motility in vitrified media with CPAs (54.4 ± 11%) was statistically higher than in media without CPAs (51.14 ± 10.6%, p < 0.05). CPAs did not affect sperm membrane integrity or large halo ratio (71.34 ± 8.47 vs. 70.38 ± 8.11 and 50.84 ± 18.92 vs. 51.98 ± 19.44, respectively). Group 2 exhibited a lower DNA fragmentation index than group 1 after vitrification (14.2 ± 8.47 vs. 12.60 ± 9.03, p = 0.021).
Conclusion: Using a pulled-glass capillary for sperm vitrification, the presence of CPAs in the vitrification medium resulted in higher progressive motility and lower DNA fragmentation index than the medium without CPAs.

Keywords: Vitrification, Sperm, DNA fragmentation, Cryoprotectant agents, Small volume.
Full-Text [PDF 1232 kb]   (419 Downloads) |   |   Full-Text (HTML)  (41 Views)  
Type of Study: Original Article | Subject: Reproductive Andrology
References
1. Hosseini A, Khalili MA, Talebi AR, Agha-Rahimi A, Ghasemi-Esmailabad S, Woodward B, et al. Cryopreservation of low number of human spermatozoa; which is better: Vapor phase or direct submerging in liquid nitrogen? Hum Fertil 2019; 22: 126-132. [DOI:10.1080/14647273.2018.1456681] [PMID]
2. Pabón D, Meseguer M, Sevillano G, Cobo A, Romero JL, Remohí J, et al. A new system of sperm cryopreservation: Evaluation of survival, motility, DNA oxidation, and mitochondrial activity. Andrology 2019; 7: 293-301. [DOI:10.1111/andr.12607] [PMID]
3. 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: 352-358. [DOI:10.1016/j.rbmo.2013.11.015] [PMID]
4. Zhu J, Jin RT, Wu LM, Johansson L, Guo TH, Liu YS, et al. Cryoprotectant-free ultra-rapid freezing of human spermatozoa in cryogenic vials. Andrologia 2014; 46: 642-649. [DOI:10.1111/and.12131] [PMID]
5. Onofre J, Baert Y, Faes K, Goossens E. Cryopreservation of testicular tissue or testicular cell suspensions: A pivotal step in fertility preservation. Hum Reprod Update 2016; 22: 744-761. [DOI:10.1093/humupd/dmw029] [PMID] [PMCID]
6. Zhao G, Fu J. Microfluidics for cryopreservation. Biotechnol Adv 2017; 35: 323-336. [DOI:10.1016/j.biotechadv.2017.01.006] [PMID] [PMCID]
7. Isachenko V, Rahimi G, Mallmann P, Sanchez R, Isachenko E. Technologies of cryoprotectant-free vitrification of human spermatozoa: Asepticity as criterion of effectiveness. Andrology 2017; 5: 1055-1063. [DOI:10.1111/andr.12414] [PMID]
8. Liu J, Tanrikut C, Wright DL, Lee GY, Toner M, Biggers JD, et al. Cryopreservation of human spermatozoa with minimal non-permeable cryoprotectant. Cryobiology 2016; 73: 162-167. [DOI:10.1016/j.cryobiol.2016.08.004] [PMID] [PMCID]
9. Schulz M, Risopatrón J, Uribe P, Isachenko E, Isachenko V, Sánchez R. Human sperm vitrification: A scientific report Mabel. Andrology 2020; 8: 1642-1650. [DOI:10.1111/andr.12847] [PMID]
10. Spis E, Bushkovskaia A, Isachenko E, Todorov P, Sanchez R, Skopets V, et al. Conventional freezing vs. cryoprotectant-free vitrification of epididymal (MESA) and testicular (TESE) spermatozoa: Three live births. Cryobiology 2019; 90: 100-102. [DOI:10.1016/j.cryobiol.2019.08.003] [PMID]
11. Chen Y, Li L, Qian Y, Xu C, Zhu Y, Huang H, et al. Small-volume vitrification for human spermatozoa in the absence of cryoprotectants by using Cryotop. Andrologia 2015; 47: 694-699. [DOI:10.1111/and.12320] [PMID]
12. Akiyama Y, Shinose M, Watanabe H, Yamada S, Kanda Y. Cryoprotectant-free cryopreservation of mammalian cells by superflash freezing. Proc Natl Acad Sci 2019; 116: 7738-7743. [DOI:10.1073/pnas.1808645116] [PMID] [PMCID]
13. Liu S, Li F. Cryopreservation of single-sperm: Where are we today? Reprod Biol Endocrinol 2020; 18: 41. [DOI:10.1186/s12958-020-00607-x] [PMID] [PMCID]
14. World Health Organization. WHO laboratory manual for the examination and processing of human semen. 5th Ed. Geneva: World Health Organization Press; 2010.
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; 46: 67-75. [DOI:10.5653/cerm.2019.46.2.67] [PMID] [PMCID]
16. Nagy ZP, Varghese AC, Agarwal A. Cryopreservation of mammalian gametes and embryos: Methods and protocols. US: Humana Press; 2017. [DOI:10.1007/978-1-4939-6828-2]
17. Widyastuti R, Lesmana R, Boediono A, Sumarsono SH. Effect of cryoprotectants on sperm vitrification. In: Hofstra R, Koibuchi N, Fucharoen S. Advance in biomolecular medicine. 1st Ed. US: CRC Press; 2017. [DOI:10.1201/9781315208619-26]
18. Le MT, Nguyen TTT, Nguyen TT, Nguyen VT, Nguyen TTA, Nguyen VQH, 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: 14-20. [DOI:10.1016/j.ejogrb.2019.01.001] [PMID]
19. O'Neill HC, Nikoloska M, Ho H, Doshi A, Maalouf W. Improved cryopreservation of spermatozoa using vitrification: Comparison of cryoprotectants and a novel device for long-term storage. J Assist Reprod Genet 2019; 36: 1713-1720. [DOI:10.1007/s10815-019-01505-x] [PMID] [PMCID]
20. Nabavinia MS, Yari A, Ghasemi-esmailabad S, Gholoobi A, Gholizadeh L, Nabi A, et al. Improvement of human sperm properties with platelet-rich plasma as a cryoprotectant supplement. Cell Tissue Bank 2023; 24: 307-315. [DOI:10.1007/s10561-022-10032-6] [PMID]
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Designed & Developed by : Yektaweb