Cryopreservation and evaluation of Akkaraman ram semen with 7-dehydrocholesterol

Abstract

The experiment was conducted to determine the in vitro results of 7-dehydrocholesterol loaded cyclodextrin compound (7-DCLC) addition to tris extenders for freezing ram semen. Three Akkaraman rams, local species in Turkey, were used. Semen samples were extended with tris containing 0 mg (control), 1.5 mg, 2.5 mg and 3.5 mg/120x106 7-DCLC and they were equilibrated to 4°C and frozen in mini straws then stored in liquid nitrogen. Straws were thawed at 37°C for 30 s in water bath for spermatological examination. Statistically significant differences were observed between 7-DCLC and control groups in terms of acrosome integrity, CASA motility, HOS test and viability after thawing (P <0.001). In all these parameters, control group had the lowest percentages. There was no statistically significant difference between groups in terms of DNA fragmentation, abnormal spermatozoa ratio, mitochondrial activation, lipid peroxidation and apoptosis level (P> 0.05). In conclusion, it can be said that adding 7-DCLC to Akkaraman ram semen improved cell membrane cryosurvival. Besides, 7-DCLC can be used in tris extender instead of cholesterol loaded cyclodextrin (CLC) compounds for sperm cryopreservation

Keywords

• Cholesterol loaded cyclodextrin
• freezing-thawing
• ram sperm
• 7-dehydrocholesterol loaded cyclodextrin

Akkaraman koç spermasının 7-dehidrokolesterol ile dondurulması ve değerlendirilmesi

Abstract

Bu çalışma, tris sulandırıcısına 7-dehidrokolesterol ile doyurulmuş siklodekstrin bileşimi (7-DCLC) eklenerek dondurmaçözdürme sonrası in vitro sonuçlarını değerlendirmek üzere yapıldı. Çalışmada yerli bir ırk olarak üç adet Akkaraman koç kullanıldı. Sperma örnekleri 0 mg (kontrol), 1.5, 2.5 ve 3.5 mg/120x106 7-DCLC içeren tris sulandırıcısı ile sulandırıldı, 4 C⁰’de alışıma bırakıldı ve mini payetlerde dondurularak sıvı azotta saklandı. Daha sonra payetler spermatolojik analizler için 37 C⁰’de 30 saniyede su banyosunda çözdürüldü. Çözüm sonu, 7-DCLC grupları ve kontrol grubu arasında CASA motilitesi, canlılık, akrozomal bütünlük ve HOS test yönünden istatiksel olarak farklılık önemli bulundu (P<0.001). Bütün bu parametrelerde kontrol gruplarında en düşük değerler tespit edildi. Anormal spermatozoa oranı, lipit peroksidasyon seviyesi, DNA hasarı, mitokondriyal aktivasyon, lipit peroksidasyon ve apoptozis seviyesi yönünden istatiksel açıdan bir farklılık bulunamadı (P>0.05). Çalışma sonucunda, 7-DCLC’nin Akkaraman koç spermasına eklenmesinin hücre membranlarını soğuktan koruduğu söylenebilir. Ayrıca, spermanın dondurulması için tris sulandırıcılarında kolesterol ile doyurulmuş siklodekstrin (CLC) bileşimi yerine 7-DCLC’nin kullanılabileceği tespit edildi

Keywords

• Çözdürme
• dondurma
• koç sperması
• kolesterol ile doyurulmuş siklodekstrin
• 7-dehidrokolesterol ile doyurulmuş siklodekstrin

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