Mikro akışkan çip, yüzdürme (swim up) ve gradient yöntemleri kullanılarak hazırlanan sperm örneklerinde DNA fragmantasyon oranlarının karşılaştırılması
Küçük Resim Yok
Tarih
2018
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Maltepe Üniversitesi Tıp Fakültesi
Erişim Hakkı
CC0 1.0 Universal
info:eu-repo/semantics/openAccess
info:eu-repo/semantics/openAccess
Özet
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<0.002). Swimup ile mikro akışkan çip yöntemleri karşılaştırıldığında DNA fragmantasyonu açısında istatistiksel olarak anlamlı bir fark izlenmedi. (p=0.583). Sonuç: Mevcut çalışmada santrifüj olmayan swim-up ve microfluidic chip yöntemlerine kıyasla gradient yöntemiyle sperm DNA fragmantasyon oranlarında artış tespit edilmiştir. Santrifüj oksidatif stresi arttırabilir ve böylece serbest radikal üretimine yol açabilir.
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<0.002). There was no statistical difference between non-centrifugal swim-up and microfluidic chip methods (p=0.583). Conclusion: Current study determined increased sperm DNA fragmentation rate by gradient method compared to non-centrifugal swim-up and microfluidic chip methods. Centrifugation may increase oxidative stress, and thus lead to free radical production.
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<0.002). There was no statistical difference between non-centrifugal swim-up and microfluidic chip methods (p=0.583). Conclusion: Current study determined increased sperm DNA fragmentation rate by gradient method compared to non-centrifugal swim-up and microfluidic chip methods. Centrifugation may increase oxidative stress, and thus lead to free radical production.
Açıklama
Anahtar Kelimeler
Sperm, DNA Fragmentasyonu, TUNEL, Sperm Yıkama, Sperm washing
Kaynak
Maltepe Tıp Dergisi / Maltepe Medical Journal
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
10
Künye
Murtaz, Tuğçe. Institute of Health Sciences, Maltepe University, Istanbul Turkey. Öztürk, Güler. Department of Physiology, Istanbul Medeniyet University School of Medicine, Istanbul Turkey. Cincik, Mehmet. Department of Histology and Embryology, Maltepe University School of Medicine, Istanbul Turkey. Selam, Belgin. Department of Obstetrics and Gynecology, Acibadem Mehmet Ali Aydinlar University, School of Medicine; Acibadem Altunizade Hospital, Unit of ART.
