Tugce MURTAZ, Guler OZTURK, Mehmet CİNCİK, Belgin SELAM

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

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

Anahtar Kelimeler:

Sperm, DNA Fragmentasyonu, TUNEL, Sperm Yıkama

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

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

Keywords:

Sperm, DNA Fragmentation, TUNEL, Sperm Washing,

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