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allied
academies
Cell Science, Stem Cell Research &
Pharmacological Regenerative Medicine
November 29-30, 2017 | Atlanta, USA
Annual Congress on
Adv cel sci tissue cul 2017 | Volume 1 Issue 2
C
ardiovascular diseases (CVDs) remain the foremost
reason of mortality and debility accounting for 31% of
deaths worldwide. Currently, regeneration of damaged
cardiac tissue with functional cardiomyocytes via stem
cell therapy represents an effective approach in CVDs
treatment. In this study, differentiation of cardiosphere
derived cells (CDCs) to cardiomyocytes was performed
in search of enhanced cardiovascular regeneration
therapeutics. Cardiac Progenitor Cells (CPCs) from rat
heart were propagated by explant culture and CDCs were
derived. Cardiac explant outgrowth cells (CEOCs) and
CDCs were characterized by Immunofluorescence, flow
cytometry and reverse transcriptase PCR. Further, CDCs
were treated with an optimized concentration of 10µM
5-Azacytidine for 24 h followed by supplementation with
10-4M ascorbic acid for 14 days. Extent of differentiation
was analyzed by immunofluorescence and quantitative real-
time PCR (qRT-PCR). In the results, flow cytometric analysis
has demonstrated that 30.23% of CEOCs were positive for
the c-kit marker, specific to CPCs. Gene expression analysis
showing high expression of
GATA4
,
Nkx2.5
and
CD90
markers suggested enhanced cardiac lineage commitment
of CDCs in comparison to CEOCs. Immunofluorescence
results confirmed that 5-Aza+AA treated CDCs expressed
cardiomyogenic markers i.e. α-sarcomeric actinin and
Nkx2.5. qRT-PCR analysis revealed relative up-regulation
of Nkx2.5,
GATA4
and
α-MHC
markers in 5-Aza+AA treated
CDCs while Wnt markers, Wnt 3a, β-catenin and cyclin D1
were down regulated. Generation of spontaneous beating
of 5-Aza+AA treated CDCs further reinforces that 5-Aza+AA
efficiently differentiated CDCs. The cardiomyogenic potential
of CDCs indicates that they can serve as an effective cellular
therapeutic as well as an ideal candidate for the treatment of
cardiovascular disorders.
e:
anu@annauniv.eduTransforming stem cell research to cardiovascular remodeling
Anuradha Dhanasekaran
and
Reddy Sailaja Mundre
Anna University, India