In vitro cardiogenesis can be initiated in human CD34+ cells.
BACKGROUND
The extensive damage that occurs in the cardiac tissue after myocardial infarct is the major concern in post infarct management. It is very well known that adult stem cells mobilized by administration of G-CSF result in homing of stem cells into the damaged myocardium. This is because of the fact that stem cells have the ability to proliferate and capacity to generate into multiple cell lineages.


METHODS
A healthy donor was selected as per the guidelines given by the institutional ethical committee and Helsinki declaration. The donor was given G-CSF 5 microg/kg/day and stem cells were harvested from the peripheral blood using Fresenius ASTec204 cell separator. The PBSC were then evaluated by immunohistochemical staining using anti-human CD34 monoclonal antibodies. The cells were then cultured in DMEM with 10% FCS for 17 weeks and in vitro cardiogenesis was initiated by adding 4 microM/l 5'Azacytidine.


RESULTS
In vitro cardiogenesis was initiated in pure CD34+ cells with 5' Azacytidine. The cells showed spontaneous beating after 24 hours of treatment and after 5 weeks, the cells connected with the adjoining cells by a myotube. In these cells, expression of myosin light chain (MLC2v) gene and GATA-4 transcription factor validated the development of cardiomyocytes.


CONCLUSIONS
It is observed that the transplantation of autologous stem cells/fetal cardiomyocytes in the heart scar tissue developed due to infarct, limited the scar expansion, and prevented post infarct heart failures. Homing process due to the transplantation of autologous stem cells is time consuming; therefore, transplantation of cardiomyocytes developed from autologous stem cells could be the future method of correcting the infracted myocardium.
Sarma
P V G K
PV
Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati 517507, India.
Subramanyam
G
G
D053687:Adult Stem Cells / Q000276:immunology*; D018952:Antigens, CD34 / Q000276:immunology*; D050991:GATA4 Transcription Factor; D016179:Granulocyte Colony-Stimulating Factor; D006801:Humans; D066298:In Vitro Techniques; D007377:Interleukin-3; D009206:Myocardium / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000276:immunology*; D013997:Time Factors
D018952:Antigens, CD34; D050991:GATA4 Transcription Factor; D007377:Interleukin-3; D016179:Granulocyte Colony-Stimulating Factor