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The GENTIL Method for Isolation of Human Adult Cardiomyocytes from Cryopreserved Tissue for Proteomic Analyses

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Familial Cardiomyopathies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2735))

Abstract

Heart failure is a serious clinical and economic health care problem, and its clinical progression is linked to pathological cardiac remodeling. Due to the heterogeneity of heart failure, lack of animal models to accurately represent advanced heart failure, and limited access to fresh human cardiac tissue, little is known regarding cell-type-specific mechanisms and context-specific functions of cardiomyocytes during disease development processes. While mass spectrometry has been increasingly applied to unravel changes in the proteome associated with cardiovascular physiology and disease, most studies have used homogenized tissue. Therefore, new studies using isolated cardiomyocytes are necessary to gain a better understanding of the intricate cell-type-specific molecular mechanisms underlying the pathophysiology of heart failure. This chapter describes the GENTIL method, which incorporates recent technological developments in sample handling, for isolation of cardiomyocytes from cryopreserved human cardiac tissues for use in proteomic analyses.

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Acknowledgments

This work was supported by the National Institutes of Health [R35-HL155460 to R.L.G.; TL1TR001437 to L.B.L], American Heart Association [20PRE35200049 to L.B.L]. Funding sources were not involved in study design, data collection, interpretation, analysis, or publication. Mass spectrometry analyses were performed using instrumentation in the CardiOmics Program in the Center for Heart and Vascular Research at UNMC.

Conflicts of Interest

R.L.G. is on the advisory board of ProtiFi and receives no compensation of any kind for this role.

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Author notes

  1. Michelle Waknitz, Linda Berg Luecke, and Roneldine Mesidor have contributed equally to this chapter

Authors and Affiliations

  1. CardiOmics Program, Center for Heart and Vascular Research; Division of Cardiovascular Medicine; and Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, NE, USA

    Michelle Waknitz, Linda Berg Luecke, Roneldine Mesidor, Melinda Wojtkiewicz, Chase Castro & Rebekah L. Gundry

  2. Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA

    Linda Berg Luecke

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  1. Michelle Waknitz
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  2. Linda Berg Luecke
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  3. Roneldine Mesidor
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  4. Melinda Wojtkiewicz
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  5. Chase Castro
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  6. Rebekah L. Gundry
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Corresponding author

Correspondence to Rebekah L. Gundry .

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Editors and Affiliations

  1. Department of Bioengineering, University of Washington, Seattle, WA, USA

    Michael Regnier

  2. Department of Bioengineering, University of Washington, Seattle, WA, USA

    Matthew Childers

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Waknitz, M., Berg Luecke, L., Mesidor, R., Wojtkiewicz, M., Castro, C., Gundry, R.L. (2024). The GENTIL Method for Isolation of Human Adult Cardiomyocytes from Cryopreserved Tissue for Proteomic Analyses. In: Regnier, M., Childers, M. (eds) Familial Cardiomyopathies. Methods in Molecular Biology, vol 2735. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3527-8_9

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  • DOI: https://doi.org/10.1007/978-1-0716-3527-8_9

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  • Publisher Name: Humana, New York, NY

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