Cardiomyocytes pp 153-160 | Cite as

Cryopreservation of Neonatal Cardiomyocytes

  • Adam C. Vandergriff
  • M. Taylor Hensley
  • Ke Cheng
Part of the Methods in Molecular Biology book series (MIMB, volume 1299)


Cardiomyocytes are frequently used for in vitro models for cardiac research. The isolation of cells is time-consuming and, due to the cells limited proliferative abilities, must be performed frequently. To reduce the time requirements and the impact on research animals, we describe a method for cryopreserving neonatal rat cardiomyocytes (NRCMs), and subsequently thawing them for use in assays.

Key words

Cardiomyocytes Cryopreservation Cardiac research 



This work was supported by funding from American Heart Association 12BGIA12040477, NC State University Chancellor’s Faculty Excellence Program, and National Natural Science Foundation of China H020381370216.


  1. 1.
    Louch WE, Sheehan KA, Wolska BM (2011) Methods in cardiomyocyte isolation, culture, and gene transfer. J Mol Cell Cardiol 51:288–298, Available at: Accessed 21 Oct 2013CrossRefPubMedCentralPubMedGoogle Scholar
  2. 2.
    Miragoli M, Salvarani N, Rohr S (2007) Myofibroblasts induce ectopic activity in cardiac tissue. Circ Res 101:755–758PubMedGoogle Scholar
  3. 3.
    Simpson P, McGrath A, Savion S (1982) Myocyte hypertrophy in neonatal rat heart cultures and its regulation by serum and by catecholamines. Circ Res 51:787–801, Available at: 1161/01.RES.51.6.787. Accessed 6 Mar 2014
  4. 4.
    Rohr S, Flückiger-Labrada R, Kucera JP (2003) Photolithographically defined deposition of attachment factors as a versatile method for patterning the growth of different cell types in culture. Pflügers Arch 446:125–32, Available at: Accessed 26 Aug 2014PubMedGoogle Scholar
  5. 5.
    Fu J-D et al (2006) Crucial role of the sarcoplasmic reticulum in the developmental regulation of Ca2+ transients and contraction in cardiomyocytes derived from embryonic stem cells. FASEB J 20:181–3, Available at: CrossRefPubMedGoogle Scholar
  6. 6.
    Liu J, Fu JD, Siu CW, Li RA (2007) Functional sarcoplasmic reticulum for calcium handling of human embryonic stem cell-derived cardiomyocytes: insights for driven maturation. Stem Cells 25:3038–44, Available at: Accessed 31 July 2014CrossRefPubMedGoogle Scholar
  7. 7.
    Knollmann BC (2013) Induced pluripotent stem cell-derived cardiomyocytes: boutique science or valuable arrhythmia model? Circ Res 112:969–976, Available at: 3667201&tool=pmcentrez&rendertype= abstract. Accessed 8 Aug 2014
  8. 8.
    Uchida T et al (2011) Optimal temperature range for low-temperature preservation of dissociated neonatal rat cardiomyocytes. Cryobiology 63:279–84, Available at: Accessed 12 Feb 2014CrossRefPubMedGoogle Scholar
  9. 9.
    Miyamura K et al (2010) Evaluation of viability of cryopreserved rat cardiac myocyte. Cryobiology 56:111–117Google Scholar
  10. 10.
    Rana P, Anson B, Engle S, Will Y (2012) Characterization of human-induced pluripotent stem cell-derived cardiomyocytes: bioenergetics and utilization in safety screening. Toxicol Sci 130:117–31, Available at: Accessed 15 July 2014CrossRefPubMedGoogle Scholar
  11. 11.
    Yokomuro H, Mickle DA, Weisel RD, Li RK (2003) Optimal conditions for heart cell cryopreservation for transplantation. Mol Cell Biochem 242:109–14, Available at: CrossRefPubMedGoogle Scholar
  12. 12.
    Simpson P, Savion S (1982) Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardial cells. Cross-striations, ultrastructure, and chronotropic response to isoproterenol. Circ Res 50:101–116, Available at: Accessed 20 Mar 2014CrossRefPubMedGoogle Scholar
  13. 13.
    Evans HJ, Goodwin RL (2007) Western array analysis of cell cycle protein changes during the hyperplastic to hypertrophic transition in heart development. Mol Cell Biochem 303:189–99, Available at: Accessed 18 Aug 2014CrossRefPubMedGoogle Scholar
  14. 14.
    Golden HB et al. (2012) Isolation of Cardiac Myocytes and Fibroblasts from Neonatal Rat Pups. In: Peng X, Antonyak M (eds) Cardiovascular Development. Methods in Molecular Biology, vol 843. Springer, New York, pp 205–214Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Adam C. Vandergriff
    • 1
    • 2
  • M. Taylor Hensley
    • 1
  • Ke Cheng
    • 2
    • 3
  1. 1.Department of Molecular Biomedical Sciences, College of Veterinary MedicineNorth Carolina State UniversityRaleighUSA
  2. 2.UNC/NCSU Joint Department of Biomedical EngineeringChapel HillUSA
  3. 3.Department of Molecular Biomedical Sciences, College of Veterinary MedicineNorth Carolina State UniversityRaleighUSA

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