Sığır oositlerinin in vitro maturasyonunun belirlenmesinde kullanılan kriterlerin etkinliğinin karşılaştırılması

Abstract

Çalışmada, mezbahadan toplanan ovaryumlardan aspire edilen toplam 200 sığır oositi kullanıldı. In vitro maturasyon, %0.6 sığır serum albumini katılmış doku kültürü vasatı içerisinde, 39°C’de, %5 CO2 atmosferinde 24 saatte gerçekleştildi. In vitro fertilizasyon işlemi ise modifiye Tyrode’nin albumin laktat piruvat vasatında aynı inkubasyon koşullarında 20 saatte gerçekleştirildi. Çalışmaya alınan oositler 10’arlı gruplar halinde işaretli petrilerde inkübe edilerek, her oosit, maturasyon sonrası kumulus ekspansiyon derecesi (yok-0, hafif-1, ileri-2), kutup hücresinin görülmesi (yok-0, var-1) ve fertilizasyonun ortaya konması (dişierkek pronukleuslar yok-0, var-1) açısından değerlendirildi. Çalışma sonucunda, in vitro maturasyon ve fertilizasyon işlemine alınan toplam 200 oositten 91’inde (%45.5) fertilizasyon saptanmıştır. Kumulus ekspansiyonunu göstermeyen 42 oositten 4’ünde fertilizasyon gözlenirken (%9.5) (p

Keywords

• İn vitro maturasyon,kumulus ekspansiyonu,kutup hücresi,oosit,sığır

The comparison of the effectiveness of the criterions used to determine the in vitro maturation of bovine oocytes

Abstract

In this study, totally 200 bovine oocytes aspirated from the slaughtered ovaries were used as the material. In vitro maturation process was carried out in bovine serum albumine added (%0.6) tissue culture media at 39 C° under 5% CO2 atmosphere for 24 hours. In vitro fertilization was carried out in modified Tyrode’s albumine lactate pyruvate media with similar conditions for 20 hours. Oocytes were placed into seperated-tagged petri dishes in ten-a-piece and every oocytes were evaluated for the cumulus expansion degrees (none-0, slight-1 and progressive-2), extursion of polar body (none-0 and existence-1) and fertilization (male and female pronuclei absence-0 and existence-1). After the fertilization process, 91 of 200 oocytes were fertilized (45.5%). Fertilization was observed in 4 of 42 unexpanded oocytes (9.5%) (p

Keywords

• Bovine,cumulus expansion,in vitro maturation,oocyte,polar body

References

1. Brackett BG, Zuelke KA (1993): Analysis of factors involved in the in vitro production of bovine embryos. Theriogenology, 39, 43-64.
2. Gordon, I (1994): The Laboratory Production of Bovine Embryos. University Press, Cambridge.
3. Gordon I, Lu KH (1990): Production of embryos in vitro and its impacts on livestock production. Theriogenology, 33, 77-87.
4. Hoshi, H (2003): In vitro production of bovine embryos and their application for embryo transfer. Theriogenology, 59, 675-685.
5. Hunter AG, Moore RM (1987): Stage-dependent effects of inhibiting ribonucleic acids and protein synthesis on meiotic maturation of bovine oocytes in vitro. J Dairy Sci, 70, 1646-1651.
6. Kim KS, Mitsumizo N, Fujita K, Utsumi K (1996): The effects of follicular fluid on in vitro maturation, oocyte fertilization and the development of bovine embryos. Theriogenology, 45, 787-799.
7. Nandi S, Ravindranatha BM, Gupta PSP, Sarma PV (2002): Timing of sequential changes in cumulus cells and first polar body extrusion during in vitro maturation of buffalo oocytes. Theriogenology, 57, 1151-1159.
8. Neglia G, Gasparrini B, di Brienza VC, Palo RD, Campanile G, Presicce GA Zicarelli L (2003): Bovine and buffalo in vitro embryo production using oocytes derived from abattoir ovaries or collected by transvaginal follicle aspiration. Theriogenology, 59, 1123-1130.

9. Nelson LD, Nelson CF (2002): Handling and culture of bovine embryos: survey of media used by 26 embryo trans- fer companies in the USA. Theriogenology, 56, 1377-1382.
10. Shamsuddin M, Larsson B, Rodriguez-Martinez H (1993): Maturation-related changes in bovine oocytes under different culture conditions. Anim Reprod Sci, 31, 49-60.
11. Sirard MA, Blondin P (1996): Oocyte maturation and IVF in cattle. Anim Reprod Sci, 42, 417-426.
12. Tanghe S, Soom AV, Mehrzad J, Maes D, Duchateau L, de Kruif A (2003): Cumulus contributions during bovine fertilization in vitro. Theriogenology, 60, 135-149.
13. Ün M (2002): Mezbahadan Toplanan Ovaryumlar-dan Elde Edilen Sığır Oositlerinin in vitro Maturasyonu ve Fertilizasyonu. Ankara Üniversitesi Sağlık Bilimleri Dok- tora Tezi, Ankara.