Characterization and comparison of mesenchymal stem cells derived from rat perirenal and periovarian adipose tissue

Abstract

Purpose: In rats, adipose tissue with white unilocular features is found in the perigonadal region. Adipose tissue contains adipocytes, adipocyte progenitor cells, mesenchymal stem cells, fibroblasts, endothelial cells, pericytes, and immune cells. Mesenchymal stem cells are multipotent stem cells and have been isolated from many tissues, including adipose tissue and connective tissue. Our aim in this study is to isolate mesenchymal stem cells from perirenal adipose tissue and periovarian adipose tissue and compare their features with each other. Materials and methods: Adipose tissues around the ovaries and kidneys of two 6-week-old Wistar Albino rats were used. Adipose tissue was cut into small pieces, and explant cell culture was established. Flow cytometry analysis was performed at the third passage (P3) for the characterization of isolated and produced mesenchymal stem cells. CD29, CD54, CD90, CD45, and MHC Class II were used as stem cell markers. mRNA expression changes of CDK6, Cyclin-D1, p21, caspase-3, caspase-8, and caspase-9 genes involved in cell cycle regulation and apoptosis were examined in cell groups. Results: Adherent cells that proliferated in explant cell culture were isolated and multiplied. In the flow cytometry analysis, it was observed that the cells expressed CD29, CD54, and CD90 markers, but did not express the cell surface antigens CD45 and MHC Class II. In the analyses performed with perirenal adipose tissue and periovarian adipose tissue-derived mesenchymal stem cells, no statistically significant difference was found in the expression of genes involved in the regulation of the cell cycle and apoptosis. Conclusion: The use of adipose tissue-derived mesenchymal stem cells from different regions, which have similar molecular properties to periovarian adipose tissue-derived mesenchymal stem cells, in ovarian failures may result in better development of follicles and the acquisition of quality oocytes.

Keywords

• Adipose tissue
• cell culture
• infertility
• ovary
• mesenchymal stem cell

Sıçan perirenal ve perioveryan adipoz doku kaynaklı mezenkimal kök hücrelerin karakterizasyonu ve birbirleriyle karşılaştırılması

Abstract

Amaç: Sıçanlarda, perigonodal bölgede beyaz unilokuler özellikte olan adipoz doku bulunur. Adipoz doku, adipositler yanında adiposit progenitör hücreler, mezenkimal kök hücreler, fibroblastlar, endotel hücreleri, perisitler ve immün hücreleri içerir. Mezenkimal kök hücreler multipotent özellikte olan kök hücrelerdir ve şimdiye kadar adipoz doku dahil olmak üzere bağ dokusu içeren birçok dokudan izole edilmiştirler. Bu çalışmadaki amacımız perirenal adipoz dokudan ve perioveryan adipoz dokudan mezenkimal kök hücreleri izole ederek özelliklerini birbirleriyle karşılaştırmaktır. Gereç ve yöntem: İki adet 6 haftalık Wistar Albino tipi sıçanın ovaryum ve böbrek çevresindeki adipoz dokuları çıkarıldı. Adipoz doku küçük parçalara ayrılarak eksplant hücre kültürü oluşturuldu. 826 İzole edilip üretilen mezenkimal kök hücrelerin karakterizasyonu için üçüncü pasajda (P3) flow sitometri analizi yapıldı. Kök hücre belirteçleri olarak CD29, CD54, CD90, CD45 ve MHC Class II kullanıldı. Hücre döngüsünün regülasyonunda ve apoptozda rol alan genlerde CDK6, Cyclin-D1, p21, caspase-3, caspase-8, caspase-9 genlerinin hücre gruplarındaki mRNA ekspresyon değişimleri incelendi. Bulgular: Eksplant hücre kültüründe prolifere olan adherent özellikteki hücreler izole edildi ve çoğaltıldı. Yapılan flow sitometri analizinde hücrelerin CD29, CD54 ve CD90 belirteçlerini eksprese ettikleri, buna karşılık hücre yüzey antijenleri olan CD45 ve MHC Class II’yi eksprese etmedikleri gözlemlenmiştir. Perirenal adipoz doku ve perioveryan adipoz doku kökenli mezenkimal kök hücrelerle yapılan analizlerde hücre döngüsünün regülasyonunda ve apoptozda rol alan genlerin ekspresyonlarında istatistiksel olarak bir farklılık bulunamadı. Sonuç: Perioveryan adipoz doku kaynaklı mezenkimal kök hücrelere benzer moleküler özelliklere sahip, farklı bölgelerdeki adipoz doku kökenli mezenkimal kök hücrelerin overyan yetmezliklerde kullanılması foliküllerin daha iyi gelişim göstermesine ve kaliteli oositlerin elde edilmesine neden olabilir.

Keywords

• Adipoz doku
• hücre kültürü
• infertilite
• ovaryum
• mezenkimal kök hücre

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