Highly enriched cardiomyocytes from human embryonic stem cells.
BACKGROUND
Current efforts to direct differentiation of human embryonic stem cells (hESC) into a particular cell lineage usually lead to a heterogeneous cell population with only a fraction of the desired cell type present. We show the generation of an essentially pure population of human cardiomyocytes from hESC using lineage selection.


METHODS
A construct comprising the murine alpha-myosin heavy chain (alpha-MHC) promoter driving the neomycin-resistance gene was introduced into hES3 cells to generate stable transgenic lines. Transgenic hESC lines were differentiated into cardiomyocytes and subjected to G418 selection. Both G418-selected and non-selected cardiomyocytes were characterized by immunocytochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The teratoma-forming potential of differentiated cells was assessed by injection of about 2 million cells into the hind limb muscle of SCID mice. Results After cardiac differentiation and antibiotic selection in a suspension culture process, more than 99% of the transgenic cells showed immunoreactivity to alpha-MHC and alpha-actinin; this enrichment efficiency was observed for independent transgenic cell lines. Quantitative RT-PCR analysis revealed high levels of enrichment for cardiac-specific messages in the selected population. Importantly, injection of selected cells into six SCID mice resulted in no apparent teratoma formation, in contrast to differentiated but non-selected controls.


CONCLUSIONS
Our results represent a significant step toward scalable production of pure human cardiomyocytes from stable, expandable hESC lines that will facilitate the development of cell therapies, safety pharmacology and drug discovery.
ES Cell International Pte Ltd, Singapore.
X Q
XQ
Xu
R
R
Zweigerdt
S Y
SY
Soo
Z X
ZX
Ngoh
S C
SC
Tham
S T
ST
Wang
R
R
Graichen
B
B
Davidson
A
A
Colman
W
W
Sun
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D004594:Electrophysiology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology; D006614:Hindlimb / Q000473:pathology; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D032383:Myocytes, Cardiac / Q000166:cytology / Q000502:physiology; D033581:Stem Cell Transplantation; D013724:Teratoma