Cardiomyogenic Gene Expression Profile of 5-Azacytidine- Treated Rat Bone Marrow Mesenchymal Stem Cells

Abstract

Cell transplantation which aims to introduce healthy myogenic cells into the myocardium of the diseased heart is an important tool in myocardial regeneration therapy. The differentiation potential of bone marrow mesenchymal stem cells BM-MSCs can be key role for this purpose. The present study aimed to evaluate cardiomyocyte gene expression profile of 5-azacytidine treated rat BM-MSCs. 5-Aza treated BM-MSCs started to form cell colonies and especially these colonies stained with desmin and cardiac troponin-T. qRT-PCR results indicate that the expression level of cardiomyocyte specific genes especially α- cardiac actin, Mef-2b, GATA-4 were increased in 5 μM 5-Aza treated BM-MSCs. According ELISA results, nanogram levels of cardiac troponin-I was measured especially 5 μM 5-Aza treated cells.

Keywords

• Bone marrow mesenchymal stem cells,5- Azacytidine,cardiomyocyte differentiation,cardiac troponin

Rat Kemik İliği Mezenşimal Kök Hücrelerinde 5-Azayctidine Uygulaması Sonucu Kardiyomiyojenik Gen İfadesi Profili

Abstract

M iyokardiyal rejenerasyon için parlak bir gelecek sunan hücresel tedavi yöntemleri hastalıklı kalbi miyojenik hücrelerle tedavi etmeyi hedeflemektedir. Kemik iliği mezenkimal kök hücrelerinin Kİ-MKH farklılaşma kapasiteleri nedeniyle miyokardiyal rejenerasyon için anahtar rol oynayabileceği düşünülmektedir. Çalışmamızda in vitro koşullarda Kİ-MKH’den kimyasal indüklemeyle kardiyomiyositlerin gen ifadesi profilinin incelenmesi amaçlanmıştır. 5-Aza uygulanan Kİ-MKH’nin kardiyomiyositlere farklılaşma süresinde, öncelikle hücreler birbirine yaklaşmaya ve hücre kümeleri oluşturmaya başlamış ve özellikle bu hücre kümelerinin desmin ve cTn-T ile immunflorasan boyamalarının pozitif olduğu görülmüştür. RT-PCR sonuçlarına göre ise farklılaşan hücrelerde kardiyak spesifik genlerden özellikle α-kardiak aktin, Mef-2b ve GATA-4 gen ifadesinin artmış olduğu saptanmıştır. ELISA sonuçlarına göre, 5 µM 5-Aza uygulanan hücrelerde nanogram seviyesinde cTn-I ölçülmüştür

Keywords

• Kemik iliği mezenkimal kök hücreleri,5- Azacytidine,hücre farklılaşması,kardiyomiyosit,kardiyak troponin

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