NKX2-5eGFP/w hESCs for isolation of human cardiac progenitors and cardiomyocytes (original) (raw)

Accession codes

Accessions

ArrayExpress

References

1. Yao, S. et al. Proc. Natl. Acad. Sci. USA 103, 6907–6912 (2006).
   Article CAS Google Scholar
2. Tomescot, A. et al. Stem Cells 133, 2200–2205 (2007).
   Article Google Scholar
3. Paige, S.L. et al. PLoS ONE 5, e11134 (2010).
   Article Google Scholar
4. Dambrot, C., Passier, R., Atsma, D. & Mummery, C.L. Biochem. J. 434, 25–35 (2011).
   Article CAS Google Scholar
5. Christoforou, N. et al. J. Clin. Invest. 118, 894–903 (2008).
   CAS PubMed PubMed Central Google Scholar
6. Satin, J. et al. Stem Cells 26, 1961–1972 (2008).
   Article CAS Google Scholar
7. Sedan, O. et al. Stem Cells 26, 3130–3138 (2008).
   Article CAS Google Scholar
8. Kehat, I. et al. J. Clin. Invest. 108, 407–414 (2001).
   Article CAS Google Scholar
9. Dick, E., Rajamohan, D., Ronksley, J. & Denning, C. Biochem. Soc. Trans. 38, 1037–1045 (2010).
   Article CAS Google Scholar
10. Yang, L. et al. Nature 453, 524–528 (2008).
    Article CAS Google Scholar
11. Bu, L. et al. Nature 460, 113–117 (2009).
    Article CAS Google Scholar
12. Laflamme, M.A. et al. Nat. Biotechnol. 25, 1015–1024 (2007).
    Article CAS Google Scholar
13. Hattori, F. et al. Nat. Methods 7, 61–66 (2010).
    Article CAS Google Scholar
14. Lints, T.J., Parsons, L.M., Hartley, L., Lyons, I. & Harvey, R.P. Development 119, 419–431 (1993).
    CAS PubMed Google Scholar
15. Kwee, L. et al. Development 121, 489–503 (1995).
    CAS PubMed Google Scholar
16. Dubois, N.C. et al. Nat. Biotechnol. (in the press).
17. Osoegawa, K. et al. Genome Res. 11, 483–496 (2001).
    Article CAS Google Scholar
18. Costa, M. et al. Nat. Protoc. 2, 792–796 (2007).
    Article CAS Google Scholar
19. Davis, R.P. et al. Nat. Protoc. 3, 1550–1558 (2008).
    Article CAS Google Scholar
20. Reubinoff, B.E., Pera, M.F., Fong, C.Y., Trounson, A. & Bongso, A. Nat. Biotechnol. 18, 399–404 (2000).
    Article CAS Google Scholar
21. Ng, E.S., Davis, R., Stanley, E.G. & Elefanty, A.G. Nat. Protoc. 3, 768–776 (2008).
    Article CAS Google Scholar
22. Ng, E.S., Davis, R.P., Azzola, L., Stanley, E.G. & Elefanty, A.G. Blood 106, 1601–1603 (2005).
    Article CAS Google Scholar
23. Burridge, P.W. et al. Stem Cells 25, 929–938 (2007).
    Article CAS Google Scholar
24. Pick, M., Azzola, L., Mossman, A., Stanley, E.G. & Elefanty, A.G. Stem Cells 25, 2206–2214 (2007).
    Article CAS Google Scholar
25. Davis, R.P. et al. Blood 111, 1876–1884 (2008).
    Article CAS Google Scholar
26. Goulburn, A.L. et al. Stem Cells 29, 462–473 (2011).
    Article CAS Google Scholar
27. Mummery, C. et al. Circulation 107, 2733–2740 (2003).
    Article CAS Google Scholar
28. Passier, R. et al. Stem Cells 23, 772–780 (2005).
    Article CAS Google Scholar
29. Prall, O.W. et al. Cell 128, 947–959 (2007).
    Article CAS Google Scholar
30. Graichen, R. et al. Differentiation 76, 357–370 (2008).
    Article CAS Google Scholar

Download references

Acknowledgements

This work was funded by the Australian Stem Cell Centre (to E.G.S., A.G.E., D.A.E., D.M.K., J.M.H. and C.W.P.), the National Health and Medical Research Council of Australia (D.A.E., grant 606586; O.W.J.P., grant 573707), National Heart Foundation (Australia) (E.G.S. and A.G.E., grant G 08M 3711; O.W.J.P., Career Development Award grant CR 08S 3958), the Netherlands Organization for Scientific Research and the Netherlands Institute of Regenerative Medicine (S.R.B. and C.L.M.), the Victorian State Government Operational Infrastructure Support and the National Health and Medical Research Council of Australia's Independent Research Institutes Infrastructure Support Scheme (O.W.J.P. and C.B.). R.P.D. is funded by a Rubicon fellowship from the Netherlands Organization for Scientific Research and the Marie Curie Co-fund Action. E.G.S. and A.G.E. receive Senior Research Fellowships and D.M.K. receives a Principal Research Fellowship from the National Health and Medical Research Council of Australia.

Author information

Author notes

1. Andrew G Elefanty and Edouard G Stanley: These authors contributed equally to this work.

Authors and Affiliations

1. Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Victoria, Australia
   David A Elliott, Katerina Koutsis, Elizabeth S Ng, Robert Jenny, Tanya Hatzistavrou, Claire E Hirst, Qing C Yu, Rhys J P Skelton, Sue Mei Lim, Xueling Li, Susan M Hawes, Adam L Goulburn, Andrew G Elefanty & Edouard G Stanley
2. Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands
   Stefan R Braam, Dorien Ward-van Oostwaard, Richard P Davis, Robert Passier & Christine L Mummery
3. Monash Institute of Pharmaceutical Science, Monash University, Victoria, Australia
   Ebba L Lagerqvist, John M Haynes & Colin W Pouton
4. The Walter and Eliza Hall Institute, Royal Melbourne Hospital, Parkville, Victoria, Australia
   Christine Biben & Owen W J Prall
5. Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia
   Ouda Khammy & David M Kaye
6. Netherlands Proteomics Institute, Utrecht, The Netherlands
   Richard P Davis & Christine L Mummery

Authors

1. David A Elliott
   You can also search for this author inPubMed Google Scholar
2. Stefan R Braam
   You can also search for this author inPubMed Google Scholar
3. Katerina Koutsis
   You can also search for this author inPubMed Google Scholar
4. Elizabeth S Ng
   You can also search for this author inPubMed Google Scholar
5. Robert Jenny
   You can also search for this author inPubMed Google Scholar
6. Ebba L Lagerqvist
   You can also search for this author inPubMed Google Scholar
7. Christine Biben
   You can also search for this author inPubMed Google Scholar
8. Tanya Hatzistavrou
   You can also search for this author inPubMed Google Scholar
9. Claire E Hirst
   You can also search for this author inPubMed Google Scholar
10. Qing C Yu
    You can also search for this author inPubMed Google Scholar
11. Rhys J P Skelton
    You can also search for this author inPubMed Google Scholar
12. Dorien Ward-van Oostwaard
    You can also search for this author inPubMed Google Scholar
13. Sue Mei Lim
    You can also search for this author inPubMed Google Scholar
14. Ouda Khammy
    You can also search for this author inPubMed Google Scholar
15. Xueling Li
    You can also search for this author inPubMed Google Scholar
16. Susan M Hawes
    You can also search for this author inPubMed Google Scholar
17. Richard P Davis
    You can also search for this author inPubMed Google Scholar
18. Adam L Goulburn
    You can also search for this author inPubMed Google Scholar
19. Robert Passier
    You can also search for this author inPubMed Google Scholar
20. Owen W J Prall
    You can also search for this author inPubMed Google Scholar
21. John M Haynes
    You can also search for this author inPubMed Google Scholar
22. Colin W Pouton
    You can also search for this author inPubMed Google Scholar
23. David M Kaye
    You can also search for this author inPubMed Google Scholar
24. Christine L Mummery
    You can also search for this author inPubMed Google Scholar
25. Andrew G Elefanty
    You can also search for this author inPubMed Google Scholar
26. Edouard G Stanley
    You can also search for this author inPubMed Google Scholar

Contributions

D.A.E., A.G.E., E.G.S. and C.L.M. designed the study. D.A.E., S.R.B., K.K., E.S.N., R.J., E.L.L., C.B., T.H., R.J.P.S., O.K., D.W.O., X.L., S.M.H., S.M.L., R.P., R.P.D., A.L.G., O.W.J.P., A.G.E., X.L., S.M.H., J.M.H., C.W.P. and D.M.K. performed and analyzed experiments. C.E.H. and Q.C.Y. performed bioinformatics analyses. D.A.E., C.L.M., A.G.E. and E.G.S. wrote the manuscript.

Corresponding authors

Correspondence toAndrew G Elefanty or Edouard G Stanley.

Ethics declarations

Competing interests

D.A.E., A.G.E. and E.G.S. have applied for a patent (US provisional USSN 61/492,099) in relation to results described in this paper.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8, Supplementary Tables 1–3 (PDF 4994 kb)

Supplementary Video 1

NKX2-5eGFP/w derived embryoid bodies express eGFP in contractile areas. Brightfield and green fluorescence of a day-14 NKX2-5eGFP/w embryoid body. This video demonstrates that contractile areas of the embryoid body express eGFP. (MOV 774 kb)

Supplementary Video 2

NKX2-5eGFP/w hESC cultured with the END2 endodermal cell line express eGFP in contractile areas. Movie shows a _Z_-dimension stack through the beating area demonstrating that eGFP is expressed with contracting clusters. (MOV 2962 kb)

Supplementary Video 3

Calcium flux across a contraction cycle. Green fluorescence (top left). Red fluorescence (top right). Brightfield (bottom left). Pseudo-colored video showing calcium flux (bottom right). (MOV 1931 kb)

Supplementary Video 4

Monolayer differentiation with NKX2-5eGFP/w hESCs. Brightfield and green fluorescence shows that beating foci are eGFP+. (MOV 936 kb)

Rights and permissions

About this article

Cite this article

Elliott, D., Braam, S., Koutsis, K. et al. NKX2-5 eGFP/w hESCs for isolation of human cardiac progenitors and cardiomyocytes.Nat Methods 8, 1037–1040 (2011). https://doi.org/10.1038/nmeth.1740

Download citation

• Received: 14 July 2011
• Accepted: 14 September 2011
• Published: 23 October 2011
• Issue Date: December 2011
• DOI: https://doi.org/10.1038/nmeth.1740