Ex vivo differentiation of human adult bone marrow stem cells into cardiomyocyte-like cells.
Bone marrow mesenchymal stem cells have been shown to transdifferentiate into cardiomyocytes after 5-azacytidine treatment or co-culturing with rodent cardiomyocytes. We investigate if adult human bone marrow stem cells can be differentiated ex vivo into cardiomyocyte-like cells (CLCs) independent of cytotoxic agents or co-culturing technique. Sternal bone marrow was collected from 16 patients undergoing coronary artery bypass surgery. Mesenchymal stem cells were differentiated in a cardiomyogenic differentiation medium containing insulin, dexamethasone, and ascorbic acid. Differentiation towards CLCs was determined by induced expression of cardiomyocyte-specific proteins. Differentiated CLCs expressed multiple structural and contractile proteins that are associated with cardiomyocytes. Thin filament associated myofibrillar proteins were detected early in the cells, with cardiac troponin I, sarcomeric tropomyosin, and cardiac titin among the first expressed. Some CLCs were found to develop into a nascent cardiomyocyte phenotype with cross-striated myofibrils characterized by alpha-actinin-positive Z bands after 4-5 passages in differentiated culture. These lineage-defined CLCs may be potentially useful for repairing damaged myocardium.
Research and Development Unit, National Heart Centre, Mistri Wing, 17 Third Hospital Ave, Singapore 168752, Singapore. Wiston_SHIM_SN@nhc.com.sg
Winston S N
WS
Shim
Shujia
S
Jiang
Philip
P
Wong
Jack
J
Tan
Yeow Leng
YL
Chua
Yong Seng
YS
Tan
Yoong Kong
YK
Sin
Chong Hee
CH
Lim
Terrance
T
Chua
Ming
M
Teh
Te Chih
TC
Liu
Eugene
E
Sim
D017667:Adipocytes / Q000378:metabolism; D000368:Aged; D001205:Ascorbic Acid / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003470:Culture Media / Q000494:pharmacology; D017931:DNA Primers / Q000737:chemistry; D003907:Dexamethasone / Q000494:pharmacology; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology; D008648:Mesoderm / Q000166:cytology; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000166:cytology; D009206:Myocardium / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D010641:Phenotype; D012313:RNA / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology; D013997:Time Factors
D003470:Culture Media; D017931:DNA Primers; D012313:RNA; D003907:Dexamethasone; D001205:Ascorbic Acid