Reproducible methodology for the isolation of mesenchymal stem cells from human umbilical cord and its potential for cardiomyocyte generation.
Mesenchymal stem cells (MSCs) are considered to be a source of stem cells in tissue regeneration and therapeutics, due to their ability to undergo proliferation and differentiation. Complications associated with bone marrow-derived MSCs has prompted researchers to explore alternative sources of MSCs. The human umbilical cord is one such source; it is easily available and its collection is non-invasive. The sources of MSCs are non-controversial and thus they are not subjected to ethical constraints, as in the case of embryonic stem cells. MSCs are multipotent stem cells and has the ability to differentiate into various cell types of the mesodermal lineage. The aim of this study was to establish a reproducible method for the isolation of MSCs from human umbilical cord, as the few methods published till date gave inconsistent results and had a mixed population of contaminating endothelial cells. In our isolation strategy, we isolated a pure population of MSCs from Wharton's jelly of the human umbilical cord, which is very rich in collagen, and we used a high concentration of collagenase enzyme in the isolation of MSCs. Extensive phenotypic characterization analysis of these cells, using flow cytometry and antibody staining methods, have shown that we were able to isolate a pure population of the mesenchymal lineage cells that is devoid of haematopoietic and endothelial cell contaminants. When these MSCs were subjected to cardiomyocyte differentiation, we observed a change in the morphological characteristics, which was accompanied by the formation of myotube structures and spontaneous beating after 21 days.
Pereira
Winston Costa
WC
R&D, LifeCell International Pvt Ltd, 26, Vandalur Kelambakkam Main Road, Keelakottaiyur, Chennai 600048, India. WinstonCosta@yahoo.com
Khushnooma
Italia
I
Madkaikar
Manisha
M
Ghosh
K
K
D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000737:chemistry*; D010641:Phenotype; D044968:Regenerative Medicine / Q000379:methods*; D015203:Reproducibility of Results; D014471:Umbilical Veins / Q000166:cytology*
D017364:Collagenases