Korhan ALTUNBAŞ, Mustafa Volkan YAPRAKÇI, Sefa ÇELIK

Isolation and Characterization of Olfactory Stem Cells from Canine Olfactory Mucosa

Olfaktorik kök hücreler nörodejeneratif hastalıkların tedavisinde büyük bir potansiyele sahiptir ve olfaktorik mukozaya kolay erişilebilirliği sayesinde hücre tedavisi için uygun bir adaydır. Bu çalışmanın amacı olfaktorik nörosfer kaynaklı hücrelere kadar farklılaştırılabilen olfaktorik kök hücrelerin izolasyonu, proliferasyonu ve karakterizasyonudur. Olfaktorik kök hücreler EGF (50 ng/ml) ve FGF (50 ng/ml) içeren serumsuz kültür vasatı içerisinde kültüre edildiğinde nörosferleri oluşturdular. Serum içeren kültür vasatında tekrar kültüre edildiklerinde olfaktorik nörosfer kaynaklı hücreleri şekillendirdiler. Gen ekspresyon analizleri OCT4, SOX2, Nanog, Nestin, ?-tubulin ve NCAM genlerinin olfaktorik kök hücrelerinde eksprese olduğunu ortaya çıkardı. Nanog, Nestin, Oct4, ? tubulin ve NCAM gen ekspresyonları nörosferlerde downregüle olurken, SOX2 geni upregüle oldu. Olfaktorik kök hücrelerin oluşturduğu nörosferlerin gen seviyeleri ile karşılaştırıldığında olfaktorik nörosfer kaynaklı hücrelerde ? tubulin ve NCAM gen ekspresyonları upregüle olurken OCT4, Nanog, Sox2 and Nestin mRNA ekspresyonları downregüle oldu. Köpeğin olfaktorik mukozası uygun alternatif bir kök hücre kaynağıdır ve köpek nörodejeneratif hastalıklarında hücre tedavisi için uygulanabilir.

Köpek Olfaktorik Mukozasindan Olfaktorik Kök Hücrelerin Izolasyonu ve Karakterizasyonu

Olfactory stem cells have great potential in the treatment of neurodegenerative diseases and they are good candidates for cell therapy due to the easy accessibility of olfactory mucosa. The main objectives of this study were isolation, proliferation and characterization of olfactory mucosa stem cells that were further differentiated into olfactory neurospheres derived cells. When grown on poly-Dlysine with a serum-free culture medium supplemented with EGF (50 ng/ml) and FGF2 (50 ng/ml), olfactory stem cells gave rise to neurospheres. When grown in serum-containing culture medium newly plated spheres gave rise to olfactory neurosphere derived cells. Gene expression analysis revealed that, OCT4, SOX2, Nanog, Nestin, ?-tubulin and NCAM were expressed in olfactory stem cells. While the mRNA expressions of Nanog, Nestin, Oct4, ?III-tubulin and NCAM were downregulated in neurospheres, the mRNA expression of SOX2 upregulated in neurospheres. According to the gene levels of neurospheres generated from olfactory stem cells, beta tubulin and NCAM gene expressions were upregulated, whereas OCT4, Nanog, Sox2 and Nestin mRNA expressions were downregulated in Olfactory neurospheres derived cells. Olfactory mucosa of canine is a suitable alternative source of stem cells and can be applied to cell therapy in neurodegenerative diseases.

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