Role of 5-azacytidine in the differentiation of rat bone marrow mesenchymal stem cells into cardiomyogenic cells
Amany Mohamed El Shawarby; Mohamed Abd Elrahman Ahmed; Mohamed Kamel Abo Golayel; Abo Zeid, Asmaa Abdelmonem;
Abstract
Introduction
Increasing attention is being paid to the use of mesenchymal stem cells (MSCs) for
treatment of human diseases such as myocardial infarction.
Aim
To study the differentiation of the bone marrow mesenchymal stem cells (BM-MSCs)
into cardiomyogenic cells using 5-azacytidine.
Materials and Methods
Forty adult male albino rats were used in this study. BM-MSCs were isolated and
cultured in a complete Dulbecco’s modified Eagle’s medium containing 1% antibiotics
and 10% fetal bovine serum (the control group). Second passaged cells were
treated with 10μmol/l 5-azacytidine for 72h. Then, the medium was removed and
kept in a 5-azacytidine-free medium for 4 weeks (the 5-azacytidine-treated group).
The adherent cells of both groups were examined using a phase-contrast microscope
and a transmission electron microscope. Expressions of cytoskeleton protein desmin and
cardiac muscle-specific cardiac troponin T were assessed by immunohistochemistry.
Results
BM-MSCs of the control group were spindle and star shaped with multiple
processes and vesicular nuclei. After adding 5-azacytidine for 1 week, the cells
showed multinucleation. On the second week, the cells formed stick-like structures.
The cells showed extensive cytoplasmic striations in the third week. Finally, in the
fourth week, the cells formed myotube-like structures. Immunohistochemical staining
of cells of the 5-azacytidine-treated group revealed a positive immune reaction for
desmin and cardiac troponin-T. Ultrastructural examination of the 5-azacytidinetreated
group revealed that the cells were elongated with central oval large nuclei.
The mitochondria were elongated with well developed cristae. There were abundant
free ribosomes and extensive dilated rough endoplasmic reticulum. Myofibrils started
to appear in the peripheral part of the cytoplasm and T-tubules appeared.
Conclusion
MSCs can be differentiated in vitro by 5-azacytidine into cardiomyogenic cells, which
are important for repairing infracted myocardium.
Keywords:
5-azacytidine, cardiac troponin T, cardiomyogenic, desmin, mesenchymal stem cells
Increasing attention is being paid to the use of mesenchymal stem cells (MSCs) for
treatment of human diseases such as myocardial infarction.
Aim
To study the differentiation of the bone marrow mesenchymal stem cells (BM-MSCs)
into cardiomyogenic cells using 5-azacytidine.
Materials and Methods
Forty adult male albino rats were used in this study. BM-MSCs were isolated and
cultured in a complete Dulbecco’s modified Eagle’s medium containing 1% antibiotics
and 10% fetal bovine serum (the control group). Second passaged cells were
treated with 10μmol/l 5-azacytidine for 72h. Then, the medium was removed and
kept in a 5-azacytidine-free medium for 4 weeks (the 5-azacytidine-treated group).
The adherent cells of both groups were examined using a phase-contrast microscope
and a transmission electron microscope. Expressions of cytoskeleton protein desmin and
cardiac muscle-specific cardiac troponin T were assessed by immunohistochemistry.
Results
BM-MSCs of the control group were spindle and star shaped with multiple
processes and vesicular nuclei. After adding 5-azacytidine for 1 week, the cells
showed multinucleation. On the second week, the cells formed stick-like structures.
The cells showed extensive cytoplasmic striations in the third week. Finally, in the
fourth week, the cells formed myotube-like structures. Immunohistochemical staining
of cells of the 5-azacytidine-treated group revealed a positive immune reaction for
desmin and cardiac troponin-T. Ultrastructural examination of the 5-azacytidinetreated
group revealed that the cells were elongated with central oval large nuclei.
The mitochondria were elongated with well developed cristae. There were abundant
free ribosomes and extensive dilated rough endoplasmic reticulum. Myofibrils started
to appear in the peripheral part of the cytoplasm and T-tubules appeared.
Conclusion
MSCs can be differentiated in vitro by 5-azacytidine into cardiomyogenic cells, which
are important for repairing infracted myocardium.
Keywords:
5-azacytidine, cardiac troponin T, cardiomyogenic, desmin, mesenchymal stem cells
Other data
| Title | Role of 5-azacytidine in the differentiation of rat bone marrow mesenchymal stem cells into cardiomyogenic cells | Authors | Amany Mohamed El Shawarby; Mohamed Abd Elrahman Ahmed; Mohamed Kamel Abo Golayel; Abo Zeid, Asmaa Abdelmonem | Issue Date | 2012 | Journal | Egyptian Journal of Histology | Volume | 35 | Issue | 1 | ISSN | 1110-0559 | DOI | 10.1097/01.ehx.0000410915.96719.4e |
Attached Files
| File | Description | Size | Format | Existing users please Login |
|---|---|---|---|---|
| Role_of_5_azacytidine_in_the_differentiation_of.4.pdf | manuscript | 807.89 kB | Adobe PDF | Request a copy |
Similar Items from Core Recommender Database
Items in Ain Shams Scholar are protected by copyright, with all rights reserved, unless otherwise indicated.