Farklı hücre senkronizasyon yöntemlerinin erken ve geç pasaj sayılarındaki sığır ve koyun hücreleri üzerindeki etkilerinin incelenmesi
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Dosyalar
Tarih
2022
Yazarlar
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Yayıncı
Tekirdağ Namık Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Donör hücrelerin hücre döngüsü aşaması, nükleer transfer embriyolarının gelişim yeteneğini etkileyen önemli bir faktördür. G0/G1 aşamaları nükleer transferde en uygun hücre döngüsü aşaması olarak kabul edilmektedir. Bu çalışmada, sığır ve koyunlardan alınan granüloza hücreleri erken (5.) ve geç (15.) pasajlarda analiz edilmektedir. Deney grupları, aktif olarak bölünen hücrelerden, 24/72 saat konfluent (%10 FBS) ve 24/72 saat serum açlığı (%0.5 FBS) uygulanan hücrelerden oluşmaktadır. Hücre döngüsü fazları, GO/G1, S ve G2/M'deki hücre yüzdeleri hücrelerin propidyum iyodür, hücre canlılığı, apoptoz/nekroz hücre yüzdeleri hücrelerin propidium iyodür ve Annexin V-FITC ile boyanması sonrasında akış sitometri cihazında belirlenmiştir. Uygulama gruplarında 24/72 saat konfluent hücrelerde G0/G1 oranı %78.69-%87.58 arasında, 24/72 saat serum açlığı uygulanan hücrelerde G0/G1 oranı %83.02 %96,16 arasında değiştiği görülerek kontrol grubundan (%68,22-76,97) yüksek bulunmuştur (P<0.05). Analiz bulgularına göre erken ve geç apoptotik hücrenin toplam oranının %1.17 %4.42 arasında, nekroz hücrelerin %1.41-%6.46 arasında değiştiği tespit edilmiştir. Koyun 15th pasaj hariç en iyi senkronizasyon serum açlığı deney gruplarından elde edilmiştir. Koyun 15th pasaj uygulama grupları arasında istatiksel olarak fark bulunmamıştır (P<0.05).
The cell cycle stage of donor cells is an important factor influencing developmental ability of nuclear transfer embryos. G0 or G1 stages are considered to be the most suitable stage for nuclear transfer. In this study, the granulosa cells from cattle and sheep were analyzed low (5th) and high (15th) passages. Experimental groups consiste actively dividing cells, contact inhibition (%10 FBS) for 24 and 72 hours, cells starvation (%0.5 FBS) for 24 and 72 hours. Cell cycle phases ratios were determined after cells were stained with propidium iodide, cell viability/apoptosis/necrosis cell ratios were determined after cells were stained with propidium iodide and Annexin V-FITC by flow cytometry In the treatment groups, it was observed that the G0/G1 rate was between %78.69-87.5 in confluent cells for 24/72 hours, and the G0/G1 rate was between %83.02-%96.16 in serum starvation cells for 24/72 hours found to be higher than the control group (%68.22-%76.97) (P<0.05). According to the analysis findings, it was determined that the total rate of early and late apoptotic cells varied between %1.17-%4.42, and necrosis cells varied between %1.41-%6.46. Except for sheep high (15th) passage, the best synchronization was obtained from serum starvation experimental groups. There was no statistical difference between the sheep higher (15th) passage application groups (P<0.05).
The cell cycle stage of donor cells is an important factor influencing developmental ability of nuclear transfer embryos. G0 or G1 stages are considered to be the most suitable stage for nuclear transfer. In this study, the granulosa cells from cattle and sheep were analyzed low (5th) and high (15th) passages. Experimental groups consiste actively dividing cells, contact inhibition (%10 FBS) for 24 and 72 hours, cells starvation (%0.5 FBS) for 24 and 72 hours. Cell cycle phases ratios were determined after cells were stained with propidium iodide, cell viability/apoptosis/necrosis cell ratios were determined after cells were stained with propidium iodide and Annexin V-FITC by flow cytometry In the treatment groups, it was observed that the G0/G1 rate was between %78.69-87.5 in confluent cells for 24/72 hours, and the G0/G1 rate was between %83.02-%96.16 in serum starvation cells for 24/72 hours found to be higher than the control group (%68.22-%76.97) (P<0.05). According to the analysis findings, it was determined that the total rate of early and late apoptotic cells varied between %1.17-%4.42, and necrosis cells varied between %1.41-%6.46. Except for sheep high (15th) passage, the best synchronization was obtained from serum starvation experimental groups. There was no statistical difference between the sheep higher (15th) passage application groups (P<0.05).
Açıklama
Anahtar Kelimeler
Ziraat, Agriculture
