Explant Culture of Ovarian Tissue

Conclusion: In the analysis, we determined that the ovarian stromal cells we isolated and produced in the culture medium expressed hematopoietic and some mesenchymal stem cell markers.

📌 3 Key Takeaways

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Objective & Hypothesis:Introduction: In this study, it was aimed to isolate, reproduce and characterize stromal cells migrating from tissues by creating ovarian tissue explant culture.

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Key Findings & Data:Materials and Methods: Explant cultures were formed from ovarian tissues of 4 week old (p repubertal) two female Wistar Albino type rats.

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Conclusion & Impact:Conclusion: In the analysis, we determined that the ovarian stromal cells we isolated and produced in the culture medium expressed hematopoietic and some mesenchymal stem cell markers.

🔬Design:Research Article

📍Scope:Comprehensive Sample

⚙️Variables:Domain Parameters

Abstract

Introduction: In this study, it was aimed to isolate, reproduce and characterize stromal cells migrating from tissues by creating ovarian tissue explant culture. Materials and Methods: Explant cultures were formed from ovarian tissues of 4 week old (p repubertal) two female Wistar Albino type rats. Then, the expression of CD29, CD54, CD90 (mesenchymal stem cell surface antigen) and CD45 (hematopoietic stem ce ll surface antigen) wa s investigated by performing flow cytometry analysis on proliferating ovarian stromal cells in the 2nd passage (P2). Results: The proliferation abilities and morphological characteristics of the cells in the culture medium were examined by serial passaging. Stromal cells migrated from the tissues and adhered to the culture dishes within the second day. They were confluent (79-80%) on the 7th day. The morphology of the ovarian stromal cells in the culture medium was observed as spindle-shaped and fibroblast-like cell aggregates when examined by phase contrast microscopy. When the proliferating stromal cells were counted under the microscope in culture dishes, 2x106 and 1.5x106 cells were observed to grow, respectively. In flow cytometry analysis of isolated ovarian stromal cells, it was determined that they expressed CD54, CD90 and CD45 surface antigens, but did not express CD29 surface antigens. Conclusion: In the analysis, we determined that the ovarian stromal cells we isolated and produced in the culture medium expressed hematopoietic and some mesenchymal stem cell markers.

Özet

Amaç: Ovaryum dokusu eksplant kültürü oluşturarak dokulardan göç eden stromal hücreleri izole etmeyi, ço ğaltmayı ve karakterize etmeyi hedefledik. Gereç ve Yöntem: 2 adet 4 haftalık (prepubertal) dişi Wistar Albino tipi sıçanların ovaryum dokularından eksplant kültürler oluşturuldu. Daha sonra prolifere olan overyan stromal hücrelere 2. pasajda (P2) flow sitometri analizi yapılarak CD29, CD54, CD90 (mezenkimal kök hücre yüzey antijeni) ve CD45 (hematopoetik kök hücre yüzey antijeni) ifadelerine bakıldı. Bulgular: Kültür ortamındaki hücrelerin proliferasyon yetenekleri ve morfolojik özellikleri seri pasajlar yapılarak incelendi. Stromal hücreler 2. günde dokulardan göç ederek kültür kaplarına yapıştılar ve çoğalıp 7. günde konflue (%70-80) oldular. Kültür ortam ındaki overyan stromal hücrelerin morfolojileri, faz kontrast mikroskobu ile incelendi ğinde iğ şeklinde ve fibroblast be nzeri hücre toplulukları olarak görüldü. Stromal hücre çoğalan kültür kaplarında mikroskop alt ında yap ılan sayımda s ırasıyla 2x106 ve 1.5x106 hücrenin ürediği gözlemlendi. İzole edilen overyan stromal hücrelerin flow sitometri analizlerinde CD54, CD90 ve CD45 yüzey antijenlerini eksprese ettikleri, CD29 yüzey antijenlerini ise eksprese etmedikleri belirlendi. Sonuç: Yapılan analizde kültür ortamında izole edip, ürettiğimiz overyan stromal hücrelerin hem hematopoetik hem de mezenkimal kök hücre belirteçlerinden bazılarını eksprese ettiklerini belirledik.

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Cite This Article

Ünal, M. S., Önder, E., Çil, N., Çoban, H. Ş., Seçme, M., Yıldırım, G., Mete, G. A. (2022). Explant Culture of Ovarian Tissue. Van Tıp Dergisi, Cilt: 29 - Sayı: 4, 421–427. https://ajindex.com/articles/655537

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