Abstract
Pediatric cardiac-derived c-kit+ cell therapies represent an innovative approach for cardiac tissue repair that have demonstrated promising improvements in recent studies and offer multiple benefits, such as easy isolation and autologous transplant. However, concerns about failure of engraftment and transient paracrine effects have thus far limited their use. To overcome these issues, an appropriate cell delivery vehicle such as a cardiac extracellular matrix (cECM) hydrogel can be utilized. This naturally derived biomaterial can support embedded cells, allowing for local diffusion of paracrine factors, and provide a healthy microenvironment for optimal cellular function. This protocol focuses on combining cardiac-derived c-kit+ cells and a cECM hydrogel to prepare a minimally invasive, dual therapeutic for in vivo delivery. We also outline a detailed method for ultrasound-guided intramyocardial injection of cell-laden hydrogels in a rodent model. Additional steps for labeling cells with a fluorescent dye for in vivo cell tracking are provided.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Tallini YN, Greene KS, Craven M, Spealman A, Breitbach M, Smith J et al (2009) C-Kit expression identifies cardiovascular precursors in the neonatal heart. Proc Natl Acad Sci U S A 106(6):1808–1813. https://doi.org/10.1073/pnas.0808920106
Agarwal U, Smith A, French KM, Boopathy AV, George A, Trac D et al (2016) Age-dependent effect of pediatric cardiac progenitor cells after juvenile heart failure. Stem Cells Transl Med 5(7):883–892. https://doi.org/10.5966/sctm.2015-0241
Trac D, Maxwell JT, Brown ME, Xu C, Davis ME (2018) Aggregation of child cardiac progenitor cells into spheres activates notch signaling and improves treatment of right ventricular heart failure. Circ Res 124(4):526–538. https://doi.org/10.1161/CIRCRESAHA.118.313845
Bejleri D, Streeter BW, Nachlas ALY, Brown ME, Gaetani R, Christman KL, Davis ME (2018) A bioprinted cardiac patch composed of cardiac-specific extracellular matrix and progenitor cells for heart repair. Adv Healthc Mater 1800672(7):1–13. https://doi.org/10.1002/adhm.201800672
Ye L, Zimmermann WH, Garry DJ, Zhang J (2013) Patching the heart: Cardiac repair from within and outside. Circ Res 113(7):922–932. https://doi.org/10.1161/CIRCRESAHA.113.300216
Feyen DAM, Gaetani R, Doevendans PA, Sluijter JPG (2016) Stem cell-based therapy: improving myocardial cell delivery. Adv Drug Deliv Rev 106:104–115. https://doi.org/10.1016/j.addr.2016.04.023
Singelyn JM, Sundaramurthy P, Johnson TD, Schup-magoffin PJ, Hu DP, Faulk DM et al (2012) Catheter-deliverable hydrogel derived from decellularized ventricular extracellular matrix increases endogenous cardiomyocytes and preserves cardiac function post-myocardial infarction. J Am Coll Cardiol 59(8). https://doi.org/10.1016/j.jacc.2011.10.888
Traverse JH, Henry TD, Dib N, Patel AN, Pepine C, Schaer GL et al (2019) First-in-man study of a cardiac extracellular matrix Hydrogel in early and late myocardial infarction patients. JACC: Basic Transl Sci 4(6):659–669. https://doi.org/10.1016/j.jacbts.2019.07.012
Ungerleider JL, Johnson TD, Hernandez MJ, Elhag DI, Braden RL, Dzieciatkowska M et al (2016) Extracellular matrix hydrogel promotes tissue remodeling, arteriogenesis, and perfusion in a Rat Hindlimb Ischemia model. JACC: Basic Transl Sci 1(1–2). https://doi.org/10.1016/j.jacbts.2016.01.009
Hernandez MJ, Gaetani R, Pieters VM, Ng NW, Chang AE, Martin TR et al (2018) Decellularized extracellular matrix hydrogels as a delivery platform for microRNA and extracellular vesicle therapeutics. Adv Ther 1800032(1):1–9. https://doi.org/10.1002/adtp.201800032
Dequach JA, Mezzano V, Miglani A, Lange S, Keller GM, Christman KL (2010) Simple and high yielding method for preparing tissue specific extracellular matrix coatings for cell culture. PLoS One 5(9):1–11. https://doi.org/10.1371/journal.pone.0013039
Singelyn JM, Dequach JA, Seif-naraghi SB, Littlefield RB, Schup-magoffin PJ, Christman KL (2010) Naturally derived myocardial matrix as an injectable scaffold for cardiac tissue engineering. Biomaterials 30(29):5409–5416. https://doi.org/10.1016/j.biomaterials.2009.06.045
French KM, Boopathy AV, DeQuach JA, Chingozha L, Lu H, Christman KL, Davis ME (2013) A naturally-derived cardiac extracellular matrix enhances cardiac progenitor cell behavior in vitro. Acta Biomater 8(12):4357–4364. https://doi.org/10.1016/j.actbio.2012.07.033.A
French KM, Davis ME (2014) Isolation and expansion of C-Kit-positive cardiac progenitor cells by magnetic cell sorting. In: Radisic M, Black LD III (eds) Cardiac tissue engineering: methods and protocols, vol 1181. Humana Press, New York, NY, pp 39–50. https://doi.org/10.1007/978-1-4939-1047-2
Ungerleider JL, Johnson TD, Rao N, Christman KL (2015) Fabrication and characterization of injectable hydrogels derived from decellularized skeletal and cardiac muscle. Methods:15–17. https://doi.org/10.1016/j.ymeth.2015.03.024
Acknowledgments
This material is based upon work that is supported by a grant from the National Institutes of Health HL146147 to MED.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Shakya, P., Brown, M.E., Davis, M.E. (2022). Encapsulation of Pediatric Cardiac-Derived C-Kit+ Cells in Cardiac Extracellular Matrix Hydrogel for Echocardiography-Directed Intramyocardial Injection in Rodents. In: Coulombe, K.L., Black III, L.D. (eds) Cardiac Tissue Engineering. Methods in Molecular Biology, vol 2485. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2261-2_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2261-2_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2260-5
Online ISBN: 978-1-0716-2261-2
eBook Packages: Springer Protocols