Dr. Ali Cihan TAŞKIN, Ahmet KOCABAY, Nilhan COŞKUN

B6D2F1 ve CB6F1 Irk Farelerde Kauda Epididimal Spermatozoa Kullanılarak İntrastoplazmik Sperm Enjeksiyonu (ICSI)

Amaç: Üreme biyoteknolojisi alanındaki çalışmalar; embriyoların uzun süre saklanması (kriyoprezervasyon), embriyo kültürü, embriyoların genom düzenlenmesi araştırmaları ve embriyo transferi gibi konular üzerinde yoğunlaşmaktadır. Üreme biyoteknolojilerinde mikromanipülasyon teknikleri, özellikle laboratuvar hayvanlarında yardımcı üreme teknolojisinin araştırıldığı çalışmalarda önemli bir yere sahiptir. Bu çalışmanın amacı, farklı fare ırklarında intrastoplazmik sperm enjeksiyonu (ICSI) ile epididimal spermatozoanın oosite enjeksiyonunu araştırmaktır. Bu çalışmada, kauda epididimal fare spermi kullanılarak yapılan ICSI uygulaması sonrasında elde edilen embriyoların in vitro gelişimi değerlendirilmiştir.Gereç ve Yöntem: Dişi fareler (8-10 hafta), gebe kısrak serum gonadotropini/insan koryonik gonadotropini (PMSG/hCG) kullanılarak süperovüle edilmiş, hCG'den ~14 saat sonra fareler sakrifiye edilerek oositler toplanmıştır. 12 haftalık erkek farenin kauda epididiminden alınan spermatozoa, aynı ırk oositle ICSI için kullanılmış ve in vitro gelişim potansiyeli değerlendirilmiştir. Son olarak tüm embriyolar 120 saat süre ile %5 CO2 ve 37°C’de kültüre edilmiştir. Bulgular: Sonuçlar, B6D2F1 (%79,31) ırkının 2 hücreli embriyo gelişiminin CB6F1 (%56,26) ırklı farelerdeki 2 hücreli embriyo gelişiminden önemli ölçüde yüksek olduğunu göstermiştir (p<0,05). Blastosist oranı B6D2F1 (%68,75) ve CB6F1 (%69,57) ırkları arasında karşılaştırılmıştır (p>0,05).Sonuç: B6D2F1 ve CB6F1 fare ırklarında, kauda epididimal sper-ma kullanılarak yapılan ICSI in vitro embriyo gelişimi için uygun bir yöntemdir. Sonuç olarak, B6D2F1 farelerde ICSI’nın başarısı, CB6F1 ırk farelere göre daha yüksektir.

Anahtar Kelimeler:

Fare, oosit, ICSI, gelişim

Intracytoplasmic Sperm Injection (ICSI) in B6D2F1 and CB6F1 Strains Mice Using Cauda Epididymal Spermatozoa

Objective: Reproductive biotechnology studies focus on the long-term storage of embryos (cryopreservation), embryo cultures, ge-nome editing of embryos and embryo transfer. Micromanipulation techniques in reproduction biotechnologies have an important role, especially in studies investigating assisted reproductive tech-nology in laboratory animals. The aim of the present study was to investigate the effect of epididymal spermatozoa injected to oo-cyte by intracytoplasmic sperm injection (ICSI) in different mice strains. In this study, we evaluated the in vitro development of post-ICSI derived embryos using cauda epididymal sperm. Material and Method: Female mice (8-10 weeks) were superovulated using pregnant mare serum gonadotropin/human chorionic gonadotropin (PMSG/hCG) and ~14h post hCG, the mice were sacrificed, and the oocytes were collected. Spermatozoa from the cau-da epididymal of a 12-week-old were used on the same strain for ICSI and the in vitro developmental potential was evaluated. Finally, the embryos were cultured for 120 hours at 5% CO2 with 37°C. Results: The results showed that the two-cell embryo of the B6D2F1 strain (79.31%) was significantly higher than the CB6F1 (56.26%) (p<0.05). While the blastocyst rate was comparable between both the B6D2F1 strain (68.75%) and CB6F1 strain (69.57%) (p>0.05).Conclusion: ICSI using cauda epididymal sperm is a suitable ap-plication for in vitro embryo development in B6D2F1 and CB6F1 strains. Finally, ICSI success of the B6D2F1 mice strains was found to be higher than CB6F1 mice strains.

Keywords:

Mouse, oocyte, ICSI, development,

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