In vitro kültürde zenginleştirilen domuz folikül ve amniyon hücrelerinden RNA ekstraksiyonu
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Date
2011-06-30
Authors
Polat, Yasemin Özen
Journal Title
Journal ISSN
Volume Title
Publisher
Uludağ Üniversitesi
Abstract
Amniyotik hücreler fetustan köken alan ve birçok farklı doku tipine dönüşebilen hücrelerdir. Bu durum hücrelerin birçok medikal uygulamada kullanılmasına olanak sağlar. Amniyotik ve foliküler sıvılar, prenatal tanıda çok etkili bir metod olan amniyosentez ile toplanmaya çalışılmıştır. Uygulamadan 48 saat sonra domuzlarda abortus görüldüğü ve toplanan hücre sayısının yeterli olmamasından dolayı, örneklerin steril şartlar altında operasyonla toplanmasına karar verilmiştir. Alınan hücreler yeterli sayı ve mitotik aktiviteye ulaşıncaya kadar hücre kültüründe geliştirilmiştir. Takibinde Trizol ile muamele edilerek hücrelerden RNA izolasyonu yapılmıştır. Amniyotik ve foliküler hücrelerin iyi kalitede üremelerine rağmen RNA izolasyonlarından alınan sonuçlar çok tatmin edici olmamıştır. RNA bütünlüğü agaroz jel elektroforezi ile kontrol edilmiştir. Elektroforez sonuçlarına göre foliküler hücrelerden elde edilen örneklere ait 28s, 18s ve 5s rRNA bantları gözlenirken, amniyotik hücre kültürlerine ait örneklerde hiç bant gözlenmemiştir. Bu sonuçlar domuza ait amniyotik ve foliküler hücrelerin in vitro ortamda üretilebildiğini göstermiştir. Ancak amniyon hücrelerine ait başarısız RNA izolasyonları açıklanamamaktadır. Amniyotik hücreler için hazırlanacak kültüre bazı zenginleştirmeler yapılabileceği yada izolasyon için daha farklı metotların kullanılabileceğini önerilebilir..
Amniotic cells are derived from fetus and they are able to differentiate into various tissue types. This allows many future medical applications. The amniotic and follicular fluids were tried to collect by amniosynthesis which is an effective method in prenatal diagnosis. Since he pigs aborted in 48 hours after the applications and the limited amount of collected samples, the collection were decided to be continued under sterile operational conditions. The cells were cultured in specific cell cultures until they reached adequate numbers of cells and mitotic activity. The cells were treated with Trizol and total RNA isolations were carried out. Although the growth of amniotic and follicular cells were in good quality, RNA isolations were not satisfactory. Integrity of the isolated RNA was verified by agarose-gel electrophoresis. The results of electrophoresis showed three clear bands of 28s, 18s, 5s rRNA from the follicular cell cultures, whereas no bands were observed from amniotic cell cultures. The results showed the proliferations of porcine follicular and amniotic cells in vitro conditions. But the unsuccessful results in RNA isolations from amniotic cells remains unclear. Different isolation techniques or utility of enriched cultures with different media can be suggested for qualified proliferations of amniotic cells.
Amniotic cells are derived from fetus and they are able to differentiate into various tissue types. This allows many future medical applications. The amniotic and follicular fluids were tried to collect by amniosynthesis which is an effective method in prenatal diagnosis. Since he pigs aborted in 48 hours after the applications and the limited amount of collected samples, the collection were decided to be continued under sterile operational conditions. The cells were cultured in specific cell cultures until they reached adequate numbers of cells and mitotic activity. The cells were treated with Trizol and total RNA isolations were carried out. Although the growth of amniotic and follicular cells were in good quality, RNA isolations were not satisfactory. Integrity of the isolated RNA was verified by agarose-gel electrophoresis. The results of electrophoresis showed three clear bands of 28s, 18s, 5s rRNA from the follicular cell cultures, whereas no bands were observed from amniotic cell cultures. The results showed the proliferations of porcine follicular and amniotic cells in vitro conditions. But the unsuccessful results in RNA isolations from amniotic cells remains unclear. Different isolation techniques or utility of enriched cultures with different media can be suggested for qualified proliferations of amniotic cells.
Description
Keywords
Amniyotik hücre, Amniotic cells, Foliküler hücre, Domuz, Follicular cells, Porcine
Citation
Özalp, G. R. vd. (2011). "In vitro kültürde zenginleştirilen domuz folikül ve amniyon hücrelerinden rna ekstraksiyonu". Uludağ Üniversitesi Veteriner Fakültesi Dergisi, 30(1), 23-28.