Influence of extenders and cooling rates on epididymal sperm of Lewis rat strain

Abstract

The objective of this study was to determine the appropriate cooling rates and extender for improving the freezability of epididymal sperm of Lewis rat strain. The epididymal sperm from Lewis rats were suspended in different freezing extenders which were HEPES buffered Tyrode’s lactate (TL-HEPES), modified Kreb’s Ringer bicarbonate (mKRB), 3% dehydrated skim milk (SM), Salamon’s Tris-citrate (TRIS), and TES/Tris (TES). The extenders contained 20% (v/v) egg yolk (EY), 0.75% equex paste (EP) and 0.1 M raffinose or 0.1 M sucrose. The sperm samples were frozen by automated freezer, using various cooling rates (10, 40, 70, and 100 °C/min). After thawing, sperm samples were examined for integrity of plasma and acrosomal integrity, and mitochondrial membrane potential (MMP). Type of extenders had significant effect on cryosurvival (p<0.05). The post-thaw highest motility (27.5%) was achieved for the sperm samples frozen in the presence of TES extender (p<0.05). As a result, although cooling rate did not significantly affect cryopreservation, the extenders affected post-thaw sperm parameters and TES considerably improved the freezability of epididymal Lewis rat sperm

Keywords

• Cooling rate
• epididymal sperm
• extenders
• freezing
• Lewis rat strain

Lewis ırkı rat sperması üzerine değişik sulandırıcı ve dondurma hızının etkisi

Abstract

Çalışmanın amacı Lewis ırkı rat sperması için, uygun dondurma hızı ve sulandırıcılar tespit ederek spermanın dondurulabilirliğinin artırılmasıdır. Ratlardan alınan epididimal sperma HEPES buffered Tyrode’s lactate (TL-HEPES), modified Kreb’s Ringer bicarbonate (mKRB), %3 süt tozu (SM), Tris-citrate (TRIS) ve Tris/TES (TES) solüsyonları ile sulandırıldı. Sulandırıcılara 20% (v/v) yumurta sarısı, 0.75% Equex Paste ve 0.1 M rafinoz yada 0.1 M sükroz ilave edildi. Sperma örnekleri kontrollü dondurucu kullanılarak değişik soğutma hızında (10, 40, 70 ve 100 °C/dk) donduruldu. Dondurma sonrası spermalar plazma, akrozom ve mitokondri membran bütünlüğü yönünden değerlendirildi. Sulandırıcı tipinin dondurmada etkili olduğu saptandı (p<0.05). Dondurma-çözüm işlemi sonrası en yüksek motilite (%27.5) TES sulandırıcısından elde edildi (p<0.05). Sonuç olarak, dondurma hızının sperm dondurulmasına önemli bir etkisi tespit edilemese de, sulandırıcıların dondurma-çözüm sonrası sperm parametrelerini etkilediği ve TES’in epididimal Lewis rat sperması dondurulabilirliğini, önemli derecede geliştirdiği saptandı

Keywords

• Dondurma
• dondurma hızı
• epididimal sperma
• sulandırıcı
• Lewis rat ırkı

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