Skip to main content
SpringerLink
Log in

Patient-Specific Induced Pluripotent Stem Cell Models: Generation and Characterization of Cardiac Cells

  • Protocol
  • First Online:
Patient-Specific Induced Pluripotent Stem Cell Models

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1353))

Abstract

The generation of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes has been of utmost interest for the study of cardiac development, cardiac disease modeling, and evaluation of cardiotoxic effects of novel candidate drugs. Several protocols have been developed to guide human stem cells toward the cardiogenic path. Pioneering work used serum to promote cardiogenesis; however, low cardiogenic throughputs, lack of chemical definition, and batch-to-batch variability of serum lots constituted a considerable impediment to the implementation of those protocols to large-scale cell biology. Further work focused on the manipulation of pathways that mouse genetics indicated to be fundamental in cardiac development to promote cardiac differentiation in stem cells. Although extremely elegant, those serum-free protocols involved the use of human recombinant cytokines that tend to be quite costly and which can also be variable between lots. The latest generation of cardiogenic protocols aimed for a more cost-effective and reproducible definition of the conditions driving cardiac differentiation, using small molecules to manipulate cardiogenic pathways overriding the need for cytokines. This chapter details methods based on currently available cardiac differentiation protocols for the generation and characterization of robust numbers of hiPSC-derived cardiomyocytes under chemically defined conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Zanella F, Lyon RC, Sheikh F (2014) Modeling heart disease in a dish: from somatic cells to disease-relevant cardiomyocytes. Trends Cardiovasc Med 24(1):32–44. doi:10.1016/j.tcm.2013.06.002

    Article  CAS  PubMed  Google Scholar 

  2. Sharma A, Wu JC, Wu SM (2013) Induced pluripotent stem cell-derived cardiomyocytes for cardiovascular disease modeling and drug screening. Stem Cell Res Ther 4(6):150. doi:10.1186/scrt380

    Article  PubMed Central  PubMed  Google Scholar 

  3. Carvajal-Vergara X, Sevilla A, D’Souza SL, Ang YS, Schaniel C, Lee DF, Yang L, Kaplan AD, Adler ED, Rozov R, Ge Y, Cohen N, Edelmann LJ, Chang B, Waghray A, Su J, Pardo S, Lichtenbelt KD, Tartaglia M, Gelb BD, Lemischka IR (2010) Patient-specific induced pluripotent stem-cell-derived models of LEOPARD syndrome. Nature 465(7299):808–812. doi:10.1038/nature09005, nature09005 [pii]

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Itzhaki I, Maizels L, Huber I, Zwi-Dantsis L, Caspi O, Winterstern A, Feldman O, Gepstein A, Arbel G, Hammerman H, Boulos M, Gepstein L (2011) Modelling the long QT syndrome with induced pluripotent stem cells. Nature 471(7337):225–229. doi:10.1038/nature09747

    Article  CAS  PubMed  Google Scholar 

  5. Moretti A, Bellin M, Welling A, Jung CB, Lam JT, Bott-Flugel L, Dorn T, Goedel A, Hohnke C, Hofmann F, Seyfarth M, Sinnecker D, Schomig A, Laugwitz KL (2010) Patient-specific induced pluripotent stem-cell models for long-QT syndrome. N Engl J Med 363(15):1397–1409. doi:10.1056/NEJMoa0908679, NEJMoa0908679 [pii]

    Article  CAS  PubMed  Google Scholar 

  6. Lahti AL, Kujala VJ, Chapman H, Koivisto AP, Pekkanen-Mattila M, Kerkela E, Hyttinen J, Kontula K, Swan H, Conklin BR, Yamanaka S, Silvennoinen O, Aalto-Setala K (2012) Model for long QT syndrome type 2 using human iPS cells demonstrates arrhythmogenic characteristics in cell culture. Dis Model Mech 5(2):220–230. doi:10.1242/dmm.008409

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Matsa E, Rajamohan D, Dick E, Young L, Mellor I, Staniforth A, Denning C (2011) Drug evaluation in cardiomyocytes derived from human induced pluripotent stem cells carrying a long QT syndrome type 2 mutation. Eur Heart J 32(8):952–962. doi:10.1093/eurheartj/ehr073

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Yazawa M, Hsueh B, Jia X, Pasca AM, Bernstein JA, Hallmayer J, Dolmetsch RE (2011) Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome. Nature 471(7337):230–234. doi:10.1038/nature09855

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Itzhaki I, Maizels L, Huber I, Gepstein A, Arbel G, Caspi O, Miller L, Belhassen B, Nof E, Glikson M, Gepstein L (2012) Modeling of catecholaminergic polymorphic ventricular tachycardia with patient-specific human-induced pluripotent stem cells. J Am Coll Cardiol 60(11):990–1000. doi:10.1016/j.jacc.2012.02.066

    Article  CAS  PubMed  Google Scholar 

  10. Fatima A, Xu G, Shao K, Papadopoulos S, Lehmann M, Arnaiz-Cot JJ, Rosa AO, Nguemo F, Matzkies M, Dittmann S, Stone SL, Linke M, Zechner U, Beyer V, Hennies HC, Rosenkranz S, Klauke B, Parwani AS, Haverkamp W, Pfitzer G, Farr M, Cleemann L, Morad M, Milting H, Hescheler J, Saric T (2011) In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells. Cell Physiol Biochem 28(4):579–592. doi:10.1159/000335753

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Jung CB, Moretti A, Mederos y Schnitzler M, Iop L, Storch U, Bellin M, Dorn T, Ruppenthal S, Pfeiffer S, Goedel A, Dirschinger RJ, Seyfarth M, Lam JT, Sinnecker D, Gudermann T, Lipp P, Laugwitz KL (2012) Dantrolene rescues arrhythmogenic RYR2 defect in a patient-specific stem cell model of catecholaminergic polymorphic ventricular tachycardia. EMBO Mol Med 4(3):180–191. doi:10.1002/emmm.201100194

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Kujala K, Paavola J, Lahti A, Larsson K, Pekkanen-Mattila M, Viitasalo M, Lahtinen AM, Toivonen L, Kontula K, Swan H, Laine M, Silvennoinen O, Aalto-Setala K (2012) Cell model of catecholaminergic polymorphic ventricular tachycardia reveals early and delayed after depolarizations. PLoS One 7(9):e44660. doi:10.1371/journal.pone.0044660

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Sun N, Yazawa M, Liu J, Han L, Sanchez-Freire V, Abilez OJ, Navarrete EG, Hu S, Wang L, Lee A, Pavlovic A, Lin S, Chen R, Hajjar RJ, Snyder MP, Dolmetsch RE, Butte MJ, Ashley EA, Longaker MT, Robbins RC, Wu JC (2012) Patient-specific induced pluripotent stem cells as a model for familial dilated cardiomyopathy. Sci Transl Med 4(130):130ra147. doi:10.1126/scitranslmed.3003552

    Article  Google Scholar 

  14. Lan F, Lee AS, Liang P, Sanchez-Freire V, Nguyen PK, Wang L, Han L, Yen M, Wang Y, Sun N, Abilez OJ, Hu S, Ebert AD, Navarrete EG, Simmons CS, Wheeler M, Pruitt B, Lewis R, Yamaguchi Y, Ashley EA, Bers DM, Robbins RC, Longaker MT, Wu JC (2013) Abnormal calcium handling properties underlie familial hypertrophic cardiomyopathy pathology in patient-specific induced pluripotent stem cells. Cell Stem Cell 12(1):101–113. doi:10.1016/j.stem.2012.10.010

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Han L, Li Y, Tchao J, Kaplan AD, Lin B, Li Y, Mich-Basso J, Lis A, Hassan N, London B, Bett GC, Tobita K, Rasmusson RL, Yang L (2014) Study familial hypertrophic cardiomyopathy using patient-specific induced pluripotent stem cells. Cardiovasc Res 104:258. doi:10.1093/cvr/cvu205

    Article  PubMed Central  PubMed  Google Scholar 

  16. Ma D, Wei H, Lu J, Ho S, Zhang G, Sun X, Oh Y, Tan SH, Ng ML, Shim W, Wong P, Liew R (2012) Generation of patient-specific induced pluripotent stem cell-derived cardiomyocytes as a cellular model of arrhythmogenic right ventricular cardiomyopathy. Eur Heart J 34:1122. doi:10.1093/eurheartj/ehs226

    Article  PubMed  Google Scholar 

  17. Kim C, Wong J, Wen J, Wang S, Wang C, Spiering S, Kan NG, Forcales S, Puri PL, Leone TC, Marine JE, Calkins H, Kelly DP, Judge DP, Chen HS (2013) Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs. Nature 494:105. doi:10.1038/nature11799

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Caspi O, Huber I, Gepstein A, Arbel G, Maizels L, Boulos M, Gepstein L (2013) Modeling of arrhythmogenic right ventricular cardiomyopathy with human induced pluripotent stem cells. Circ Cardiovasc Genet 6(6):557–568. doi:10.1161/CIRCGENETICS.113.000188

    Article  CAS  PubMed  Google Scholar 

  19. Davis RP, Casini S, van den Berg CW, Hoekstra M, Remme CA, Dambrot C, Salvatori D, Oostwaard DW, Wilde AA, Bezzina CR, Verkerk AO, Freund C, Mummery CL (2012) Cardiomyocytes derived from pluripotent stem cells recapitulate electrophysiological characteristics of an overlap syndrome of cardiac sodium channel disease. Circulation 125(25):3079–3091. doi:10.1161/CIRCULATIONAHA.111.066092

    Article  PubMed  Google Scholar 

  20. Lian X, Zhang J, Azarin SM, Zhu K, Hazeltine LB, Bao X, Hsiao C, Kamp TJ, Palecek SP (2013) Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/beta-catenin signaling under fully defined conditions. Nat Protoc 8(1):162–175. doi:10.1038/nprot.2012.150

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Burridge PW, Matsa E, Shukla P, Lin ZC, Churko JM, Ebert AD, Lan F, Diecke S, Huber B, Mordwinkin NM, Plews JR, Abilez OJ, Cui B, Gold JD, Wu JC (2014) Chemically defined generation of human cardiomyocytes. Nat Methods 11(8):855–860. doi:10.1038/nmeth.2999

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Chen G, Gulbranson DR, Hou Z, Bolin JM, Ruotti V, Probasco MD, Smuga-Otto K, Howden SE, Diol NR, Propson NE, Wagner R, Lee GO, Antosiewicz-Bourget J, Teng JM, Thomson JA (2011) Chemically defined conditions for human iPSC derivation and culture. Nat Methods 8(5):424–429. doi:10.1038/nmeth.1593

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Lian X, Hsiao C, Wilson G, Zhu K, Hazeltine LB, Azarin SM, Raval KK, Zhang J, Kamp TJ, Palecek SP (2012) Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A 109(27):E1848–E1857. doi:10.1073/pnas.1200250109

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Lian X, Zhang J, Zhu K, Kamp TJ, Palecek SP (2013) Insulin inhibits cardiac mesoderm, not mesendoderm, formation during cardiac differentiation of human pluripotent stem cells and modulation of canonical Wnt signaling can rescue this inhibition. Stem Cells 31(3):447–457. doi:10.1002/stem.1289

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

F.Z. is a current recipient of the American Heart Association Post-doctoral Fellowship. F.S. is supported by grants from the National Institutes of Health, American Heart Association and Saving tiny Heart Society grants.

Author information

Authors and Affiliations

  1. Cardiology Division, Department of Medicine, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0613C, USA

    Fabian Zanella & Farah Sheikh

Authors
  1. Fabian Zanella
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Farah Sheikh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Farah Sheikh .

Editor information

Editors and Affiliations

  1. Lunenfeld-Tanenbaum Research Institute, Toronto, Ontario, Canada

    Andras Nagy

  2. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Zanella, F., Sheikh, F. (2014). Patient-Specific Induced Pluripotent Stem Cell Models: Generation and Characterization of Cardiac Cells. In: Nagy, A., Turksen, K. (eds) Patient-Specific Induced Pluripotent Stem Cell Models. Methods in Molecular Biology, vol 1353. Humana Press, New York, NY. https://doi.org/10.1007/7651_2014_172

Download citation

  • DOI: https://doi.org/10.1007/7651_2014_172

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3033-3

  • Online ISBN: 978-1-4939-3034-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation