BibTex RIS Kaynak Göster

Comparison of DNA fragmentation rates in sperm samples prepared by microfluidic chip, swim-up and gradient methods

Yıl 2018, Cilt: 10 Sayı: 2, 49 - 53, 01.08.2018

Öz

Aim: Sperm preparation methods significantly effect the results of assisted reproduction techniques. Sperm washing removes cells potentially harmful to fertilization and obtains more motile and morphologically better sperm. Conventional sperm washing techniques, include centrifugal, non-centrifugal swim-up and centrifugal gradient methods. Sperm DNA fragmentation is one of the factors suggested for male infertility. Sperm washing methods may effect DNA fragmentation rate. The aim of the current study is to compare sperm DNA fragmentation rates between conventional sperm washing methods and relatively new microfluidic chip method. Methods: Study group consisted of 14 volunteer men. Every sperm sample was allocated and washed separately in 3 categories as; gradient, non-centrifugal swim-up and finally microfluidic chip methods. Sperm DNA fragmentation rates in each category were compared by TUNEL Terminal deoxynucleotidyl transferase dUTP nick end labeling test. Results: Ratio of TUNEL positive sperm with DNA fragmentation were 12%, 4% and 4,5% by gradient, non-centrifugal swim-up and microfluidic chip methods, respectively. Sperm DNA fragmentation ratio by gradient method statistically significantly increased compared to those with non-centrifugal swim-up and microfluidic chip methods p

Kaynakça

  • 1. Mosher WD, Pratt, WF. Fecundity and infertility in the United States: incidence and trends. Fertil Steril 1991;56:192-193.
  • 2. Thonneau, P, Marchand, S, Tallec A, Ferial ML, Ducot B, Lansac J. Incidence and main causes of infertility in a resident population (1, 850, 000) of three French regions (1988-1989). Hum Reprod 1991;6:811- 816.
  • 3. Host E, Lindenberg S, Smidt-Jensen S. The role of DNA strand breaks in human spermatozoa used for IVF and ICSI. Acta Obstet Gynecol Scand 2001;79:559- 563.
  • 4. Zini, A, Bielecki R, Phang D, Zenzes MT. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril 2001;75:674- 677.
  • 5. 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.
  • 6. Aitken RJ, Gordon E, Harkiss D, et al. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod 1998;59:1037-1046.
  • 7. Zini A, Finelli A, Phang D, Jarvi K. Influence of semen processing technique on human sperm DNA integrity, Urology, 2000;56:1081-1084.
  • 8. Aitken, RJ., Clarkson, JS. Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques. J Androl. 1988; 9:367-376.
  • 9. Russo A, Troncoso N, Sanchez F, Garbarino JA, Vanella A. Propolis protects human spermatozoa from DNA damage caused by benzopyrene and exogenous reactive oxygen species. Life Sciences 2006;78:1401- 1406.
  • 10. de Lamirande E, Gagnon C. Human sperm hyperactivation and capacitation as parts of an oxidative process. Free Radic Biol Med 1993;14:157- 166.
  • 11. Pasqualotto FF, Sharma RK, Nelson DR, Thomas AJ, Agarwal A. Relationship between oxidative stress, semen characteristics, and clinical diagnosis in men undergoing infertility investigation. Fertil Steril 2000;73:459-464.
  • 12. Sharma RK, Pasqualotto AE, Nelson DR, Thomas AJ Jr, Agarwal A. Relationship between seminal white blood cell counts and oxidative stress in men treated at an infertility clinic. J Androl 2001;22:575- 583
  • 13. de Lamirande E, Gagnon C. Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects Hum Reprod 1995;10 Suppl 1:15-21.
  • 14. Leclerc P, de Lamirande E, Gagnon C. Interaction between Ca2+, cyclic 3′,5′ adenosine monophosphate, the superoxide anion, and tyrosine phosphorylation pathways in the regulation of human sperm capacitation. J Androl 1998;19:434-443.
  • 15. Aitken RJ, Gordon E, Harkiss D, et al. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod 1998;59:1037-1046.
  • 16. de Lamirande, E., Gagnon, C. A positive role for the superoxide anion in triggering hyperactivation and capacitation of human spermatozoa. Int J Androl 1993b ;16:21-25.
  • 17. de Lamirande, E., Gagnon, C. Human sperm hyperactivation in whole semen and its association with low superoxide scavenging capacity in seminal plasma. Fertil Steril 1993c ;59: 1291-1295.
  • 18. Zini A, Libman J. Sperm DNA damage; clinical significance in the era of assisted reproduction. CMAJ 2006;175:495-500.
  • 19. Asghar W, Velasco V, Kingsley JL, et al. Selection of functional human sperm with higher DNA integrity and fewer reactive oxygen species. Adv Healthc Mater 2014;3:1671-1679.

Mikro akışkan çip, yüzdürme swim up ve gradient yöntemleri kullanılarak hazırlanan sperm örneklerinde DNA fragmantasyon oranlarının karşılaştırılması

Yıl 2018, Cilt: 10 Sayı: 2, 49 - 53, 01.08.2018

Öz

Amaç: Sperm hazırlama yöntemleri, yardımlı üreme tekniklerinin sonuçlarını önemli ölçüde etkiler. Sperm yıkama işlemi, fertilizasyon için zararlı olabilecek hücrelerin ayrıştırır ve daha hareketli ve morfolojik olarak iyi spermlerin elde edilmesini sağlar. Geleneksel sperm yıkama teknikleri, santrifüjlü ve santrifüjsüz swim-up ve santrifüjlü gradient yöntemleridir. Sperm DNA fragmantasyonu, erkek infertilitesi için önerilen faktörlerden biridir. Sperm yıkama yöntemleri DNA fragmantasyonu oranlarını etkileyebilir. Bu çalışmanın amacı, geleneksel sperm yıkama yöntemleri ile yeni bir yöntem olan mikro akışkan çip yöntemi arasında DNA hasarı oranlarını kıyaslamaktır. Gereç ve Yöntem: Çalışma grubu 14 gönüllü erkekten oluşmaktadır. Alınan her semen örneği 3’e ayrılarak gradient, santrifüjsüz swim up ve mikro akışkan çip yöntemleriyle ayrı ayrı yıkandı. Her gruptaki sperm DNA fragmantasyon oranları TUNEL terminal deoksinükleotidil transferaz-aracılı dUTP işaretleme testi ile karşılaştırıldı. Bulgular: DNA fragmantasyonlu TUNEL pozitif sperm oranları, gradient, santrifüjsüz swim up ve mikro akışkan çip yöntemleri için sırasıyla %12, %4 ve %4,5 izlendi. Gradient yöntemi ile sperm DNA fragmantasyon oranları, diğer iki yöntemle karşılaştırıldığında istatistiksel olarak anlamlı yüksek bulundu p

Kaynakça

  • 1. Mosher WD, Pratt, WF. Fecundity and infertility in the United States: incidence and trends. Fertil Steril 1991;56:192-193.
  • 2. Thonneau, P, Marchand, S, Tallec A, Ferial ML, Ducot B, Lansac J. Incidence and main causes of infertility in a resident population (1, 850, 000) of three French regions (1988-1989). Hum Reprod 1991;6:811- 816.
  • 3. Host E, Lindenberg S, Smidt-Jensen S. The role of DNA strand breaks in human spermatozoa used for IVF and ICSI. Acta Obstet Gynecol Scand 2001;79:559- 563.
  • 4. Zini, A, Bielecki R, Phang D, Zenzes MT. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril 2001;75:674- 677.
  • 5. 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.
  • 6. Aitken RJ, Gordon E, Harkiss D, et al. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod 1998;59:1037-1046.
  • 7. Zini A, Finelli A, Phang D, Jarvi K. Influence of semen processing technique on human sperm DNA integrity, Urology, 2000;56:1081-1084.
  • 8. Aitken, RJ., Clarkson, JS. Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques. J Androl. 1988; 9:367-376.
  • 9. Russo A, Troncoso N, Sanchez F, Garbarino JA, Vanella A. Propolis protects human spermatozoa from DNA damage caused by benzopyrene and exogenous reactive oxygen species. Life Sciences 2006;78:1401- 1406.
  • 10. de Lamirande E, Gagnon C. Human sperm hyperactivation and capacitation as parts of an oxidative process. Free Radic Biol Med 1993;14:157- 166.
  • 11. Pasqualotto FF, Sharma RK, Nelson DR, Thomas AJ, Agarwal A. Relationship between oxidative stress, semen characteristics, and clinical diagnosis in men undergoing infertility investigation. Fertil Steril 2000;73:459-464.
  • 12. Sharma RK, Pasqualotto AE, Nelson DR, Thomas AJ Jr, Agarwal A. Relationship between seminal white blood cell counts and oxidative stress in men treated at an infertility clinic. J Androl 2001;22:575- 583
  • 13. de Lamirande E, Gagnon C. Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects Hum Reprod 1995;10 Suppl 1:15-21.
  • 14. Leclerc P, de Lamirande E, Gagnon C. Interaction between Ca2+, cyclic 3′,5′ adenosine monophosphate, the superoxide anion, and tyrosine phosphorylation pathways in the regulation of human sperm capacitation. J Androl 1998;19:434-443.
  • 15. Aitken RJ, Gordon E, Harkiss D, et al. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod 1998;59:1037-1046.
  • 16. de Lamirande, E., Gagnon, C. A positive role for the superoxide anion in triggering hyperactivation and capacitation of human spermatozoa. Int J Androl 1993b ;16:21-25.
  • 17. de Lamirande, E., Gagnon, C. Human sperm hyperactivation in whole semen and its association with low superoxide scavenging capacity in seminal plasma. Fertil Steril 1993c ;59: 1291-1295.
  • 18. Zini A, Libman J. Sperm DNA damage; clinical significance in the era of assisted reproduction. CMAJ 2006;175:495-500.
  • 19. Asghar W, Velasco V, Kingsley JL, et al. Selection of functional human sperm with higher DNA integrity and fewer reactive oxygen species. Adv Healthc Mater 2014;3:1671-1679.
Toplam 19 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Araştırma Makalesi
Yazarlar

Tugce Murtaz Bu kişi benim

Guler Ozturk Bu kişi benim

Mehmet Cincik Bu kişi benim

Belgin Selam Bu kişi benim

Yayımlanma Tarihi 1 Ağustos 2018
Yayımlandığı Sayı Yıl 2018 Cilt: 10 Sayı: 2

Kaynak Göster

Vancouver Murtaz T, Ozturk G, Cincik M, Selam B. Mikro akışkan çip, yüzdürme swim up ve gradient yöntemleri kullanılarak hazırlanan sperm örneklerinde DNA fragmantasyon oranlarının karşılaştırılması. Maltepe tıp derg. 2018;10(2):49-53.