Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial

Reproductive Biology and Endocrinology

Muhammad Abu-Elmagd¹  Adel M Abuzenadah³  Ashok Agarwal²  Rakesh Sharma²  Helena Malvezzi² 

[1] Zoology Department, Faculty of Science, Minia University, Minia, Egypt;Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Cleveland, OH, USA;KACST Technology Innovation Center in Personalized Medicine at King AbdulAziz University, Jeddah, Saudi Arabia

关键词: DNA fragmentation;    Density gradient centrifugation;    Reproductive potential;    Sperm processing;    Sperm motility;   

Others  :  1132323
DOI  :  10.1186/1477-7827-12-121

 received in 2014-09-09, accepted in 2014-11-20,  发布年份 2014

【 摘 要 】

Background

Density gradient is the preferred technique for sperm processing for ART. However, no study has examined sperm quality using different processing media simultaneously and under identical conditions. Therefore, we evaluated semen quality following sperm preparation by three commonly used commercially available density gradient media in a well-designed controlled trial.

Methods

We obtained semen samples from 20 healthy volunteers. Percent motility, total motile sperm (TMS), % recovery and DNA damage were assessed before and after separation in three different sperm density gradient media-PureCeption, ISolate and SpermGrad-125.

Results

Percent motility was higher in the ISolate (81.4% ± 6.6%) and SpermGrad-125 samples (85.7% ± 8.0%) (P < 0.0001) than in the PureCeption samples (62.5% ± 13.2%) (P = 0.07). TMS was higher in the PureCeption(TM) and ISolate samples (14.2% ± 15.9% and 15.8% ± 18.2%) than in those prepared with SpermGrad-125 (10.6% ± 19.7%) (P < 0.0001). Percent recovery was significantly higher in the PureCeption(TM) and ISolate samples (45.3% and 48.9%) than in the SpermGrad-125(TM) samples (30.8%) (P < 0.01). DNA fragmentation was comparable across the three gradients (PureCeption = 8.8% ± 4.7%; ISolate = 7.2 ± 5.2% and SpermGrad-125 = 11.2% ± 7.4%).

Conclusions

Three different density gradient processing media PureCeption, ISolate, and SpermGrad-125 were examined for their effects on sperm quality. Sperm processed by ISolate and Sperm Grad 125 had better motility and TMS after processing. The extent of DNA damage was comparable in all three gradients.

【 授权许可】

   
2014 Malvezzi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Vanage GR, Gopalkrishnan K, Sheth AR: Effect of antibodies to human seminal plasma inhibin on spermatogenesis and sperm agglutination in adult male rats. Mol Reprod Dev 1990, 25:227-236.
• [2]Levay PF, Fourie FR, Meintjes J: The effect of seminal plasma on human sperm-zona pellucida binding. Hum Reprod 1995, 10:2590-2594.
• [3]Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ Jr, Agarwal A: Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum Reprod 2001, 16:1922-1930.
• [4]Allamaneni SS, Agarwal A, Rama S, Ranganathan P, Sharma RK: Comparative study on density gradients and swim-up preparation techniques utilizing neat and cryopreserved spermatazoa. Asian J Androl 2005, 7:86-92.
• [5]Beydola T, Sharma RK, Lee W, Agarwal A: Sperm preparation and selection techniques. In Male Infertility Practice. Edited by Rizk B, Aziz N, Agarwal A. New Delhi: Jaypee Brothers Medical Publishers; 2013:244-251.
• [6]Boomsma CM, Heineman MJ, Cohlen BJ, Farquhar C: Semen preparation techniques for intrauterine insemination. Cochrane Database Syst Rev 2004, 3:CD004507.
• [7]Jayaraman V, Upadhya D, Narayan PK, Adiga SK: Sperm processing by swim-up and density gradient is effective in elimination of sperm with DNA damage. J Assist Reprod Genet 2012, 29:557-563.
• [8]Zini A, Nam RK, Mak V, Phang D, Jarvi K: Influence of initial semen quality on the integrity of human sperm DNA following semen processing. Fertil Steril 2000, 74:824-827.
• [9]Zini A, Finelli A, Phang D, Jarvi K: Influence of semen processing technique on human sperm DNA integrity. Urology 2000, 56:1081-1084.
• [10]Bungum M, Spanò M, Humaidan P, Eleuteri P, Rescia M, Giwercman A: Sperm chromatin structure assay parameters measured after density gradient centrifugation are not predictive for the outcome of ART. Hum Reprod 2008, 23:4-10.
• [11]Brahem S, Mehdi M, Landolsi H, Mougou S, Elghezal H, Saad A: Semen parameters and sperm DNA fragmentation as causes of recurrent pregnancy loss. Urology 2011, 78:792-796.
• [12]Brahem S, Mehdi M, Elghezal H, Saad A: Semen processing by density gradient centrifugation is useful in selecting sperm with higher double-strand DNA integrity. Andrologia 2011, 43:196-202.
• [13]Enciso M, Iglesias M, Galán I, Sarasa J, Gosálvez A, Gosálvez J: The ability of sperm selection techniques to remove single- or double-strand DNA damage. Asian J Androl 2011, 13:764-768.
• [14]Lopes S, Sun JG, Jurisicova A, Meriano J, Casper RF: Sperm deoxyribonucleic acid fragmentation is increased in poor-quality semen samples and correlates with failed fertilization in intracytoplasmic sperm injection. Fertil Steril 1998, 69:528-532.
• [15]Muratori M, Piomboni P, Baldi E, Filimberti E, Pecchioli P, Moretti E, Gambera L, Baccetti B, Biagiotti R, Forti G, Maggi M: Functional and ultrastructural features of DNA-fragmented human sperm. J Androl 2000, 21:903-912.
• [16]Sharma RK, Sabanegh E, Mahfouz R, Gupta S, Thiyagarajan A, Agarwal A: TUNEL as a test for sperm DNA damage in the evaluation of male infertility. Urology 2010, 76:1380-1386.
• [17]Sharma RK, Agarwal A: Laboratory evaluation of sperm chromatin: TUNEL assay. In Sperm Chromatin: Biological and Clinical Applications in Male Infertility and Assisted Reproduction. Edited by Zini A, Agarwal A. New York: Springer Science + Business Media; 2011:201-215.
• [18]Mahfouz R, Sharma R, Thiyagarajan A, Kale V, Gupta S, Sabanegh E, Agarwal A: Semen characteristics and sperm DNA fragmentation in infertile men with low and high levels of seminal reactive oxygen species. Fertil Steril 2010, 94:2141-2146.
• [19]Agarwal A, Varghese AC, Sharma RK: Markers of oxidative stress and sperm chromatin integrity. Methods Mol Biol 2009, 590:377-402.
• [20]Makker K, Agarwal A, Sharma R: Oxidative stress & male infertility. Indian J Med Res 2009, 129:357-367.
• [21]Abd-Elmoaty MA, Saleh R, Sharma R, Agarwal A: Increased levels of oxidants and reduced antioxidants in semen of infertile men with varicocele. Fertil Steril 2010, 94:1531-1534.
• [22]Benchaib M, Ajina M, Lornage J, Niveleau A, Durand P, Guérin JF: Quantitation by image analysis of global DNA methylation in human spermatozoa and its prognostic value in in vitro fertilization: a preliminary study. Fertil Steril 2003, 80:947-953.
• [23]Virant-Klun I, Tomazevic T, Meden-Vrtovec H: Sperm single-stranded DNA, detected by acridine orange staining, reduces fertilization and quality of ICSI-derived embryos. J Assist Reprod Genet 2002, 19:319-328.
• [24]Sun JG, Jurisicova A, Casper RF: Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol Reprod 1997, 56:602-607.
• [25]Larson KL, DeJonge CJ, Barnes AM, Jost LK, Evenson DP: Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Hum Reprod 2000, 15:1717-1722.
• [26]Lewis SE, Aitken RJ: DNA damage to spermatozoa has impacts on fertilization and pregnancy. Cell Tissue Res 2005, 322:33-41.
• [27]Chiamchanya C, Kaewnoonual N, Visutakul P, Manochantr S, Chaiya J: Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters. Asian J Androl 2010, 12:271-277.
• [28]World Health Organization: WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th edition. Geneva, Switzerland;
• [29]Sharma R, Masaki J, Agarwal A: Sperm DNA fragmentation analysis using the TUNEL assay. Methods Mol Biol 2013, 927:121-136.
• [30]Chi HJ, Kim JH, Ryu CS, Lee JY, Park JS, Chung DY, Choi SY, Kim MH, Chun EK, Roh SI: Protective effect of antioxidant supplementation in sperm-preparation medium against oxidative stress in human spermatozoa. Hum Reprod 2008, 23:1023-1028.
• [31]Benchaib M, Lornage J, Mazoyer C, Lejeune H, Salle B, François Guerin J: Sperm deoxyribonucleic acid fragmentation as a prognostic indicator of assisted reproductive technology outcome. Fertil Steril 2007, 87:93-100.
• [32]Li Z, Zhou Y, Liu R, Lin H, Liu W, Xiao W, Lin Q: Effects of semen processing on the generation of reactive oxygen species and mitochondrial membrane potential of human spermatozoa. Andrologia 2012, 44:157-163.
• [33]Marchesi DE, Biederman H, Ferrara S, Hershlag A, Feng HL: The effect of semen processing on sperm DNA integrity: comparison of two techniques using the novel Toluidine Blue assay. Eur J Obstet Gynecol Reprod Biol 2010, 151:176-180.
• [34]Zhang XD, Chen MY, Gao Y, Han W, Liu DY, Huang GN: The effects of different sperm preparation methods and incubation time on the sperm DNA fragmentation. Hum Fertil (Camb) 2011, 14:187-191.
• [35]Stevanato J, Bertolla RP, Barradas V, Spaine DM, Cedenho AP, Ortiz V: Semen processing by density gradient centrifugation does not improve sperm apoptotic deoxyribonucleic acid fragmentation rates. Fertil Steril 2008, 90:889-890.
• [36]Lewis SE, Simon L: Clinical implications of sperm DNA damage. Hum Fertil (Camb) 2010, 13:201-207.
• [37]Jiang H, He RB, Wang CL, Zhu J: The relationship of sperm DNA fragmentation index with the outcomes of in-vitro fertilisation-embryo transfer and intracytoplasmic sperm injection. J Obstet Gynaecol 2011, 31:636-639.