Generation of Self-Induced Myocardial Ischemia in Large-Sized Cardiac Spheroids without Alteration of Environmental Conditions Recreates Fibrotic Remodeling and Tissue Stiffening Revealed by Constriction Assays.

Felipe Prósper, Gregorio Rábago, Miguel A Martínez, Ignacio Ochoa, Margarida Serra, Arantxa González, Pilar Montero-Calle, Manuel M Mazo, Laura Paz-Artigas, Sandra González-Lana, Nicolás Polo, Pedro Vicente, Jesús Ciriza

Journal: ACS biomaterials science & engineering 2024;10(2):987-997

PMID: 38234159

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Abstract

A combination of human-induced pluripotent stem cells (hiPSCs) and 3D microtissue culture techniques allows the generation of models that recapitulate the cardiac microenvironment for preclinical research of new treatments. In particular, spheroids represent the simplest approach to culture cells in 3D and generate gradients of cellular access to the media, mimicking the effects of an ischemic event. However, previous models required incubation under low oxygen conditions or deprived nutrient media to recreate ischemia. Here, we describe the generation of large spheroids (i.e., larger than 500 μm diameter) that self-induce an ischemic core. Spheroids were generated by coculture of cardiomyocytes derived from hiPSCs (hiPSC-CMs) and primary human cardiac fibroblast (hCF). In the proper medium, cells formed aggregates that generated an ischemic core 2 days after seeding. Spheroids also showed spontaneous cellular reorganization after 10 days, with hiPSC-CMs located at the center and surrounded by hCFs. This led to an increase in microtissue stiffness, characterized by the implementation of a constriction assay. All in all, these phenomena are hints of the fibrotic tissue remodeling secondary to a cardiac ischemic event, thus demonstrating the suitability of these spheroids for the modeling of human cardiac ischemia and its potential application for new treatments and drug research.

Address: Tissue Microenvironment (TME) Lab, Aragón Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza 50018, Spain.; Institute for Health Research Aragón (IIS Aragón), Zaragoza 50009, Spain.; Tissue Microenvironment (TME) Lab, Aragón Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza 50018, Spain.; BEONCHIP S.L., CEMINEM, Campus Río Ebro, Zaragoza 50018, Spain.; Tissue Microenvironment (TME) Lab, Aragón Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza 50018, Spain.; Instituto de Biologia Experimental e Tecnológica (iBET), Oeiras 2780-157, Portugal.; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras 2780-157, Portugal.; Cardiology and Cardiac Surgery Department, Clínica Universidad de Navarra, Pamplona 31009, Spain.; Tissue Microenvironment (TME) Lab, Aragón Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza 50018, Spain.; CIBER-BBN, ISCIII, Zaragoza 50018, Spain.; Regenerative Medicine Program, Cima Universidad de Navarra, and Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona 31008, Spain.; Hematology and Cell Therapy, Clínica Universidad de Navarra, and Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona 31008, Spain.; CIBERONC, Instituto de Salud Carlos III, Madrid 28029, Spain.; Regenerative Medicine Program, Cima Universidad de Navarra, and Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona 31008, Spain.; Hematology and Cell Therapy, Clínica Universidad de Navarra, and Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona 31008, Spain.; Tissue Microenvironment (TME) Lab, Aragón Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza 50018, Spain.; Program of Cardiovascular Diseases, CIMA Universidad de Navarra, and Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona 31008, Spain.; CIBERCV, Instituto de Salud Carlos III, Madrid 28029, Spain.; Tissue Microenvironment (TME) Lab, Aragón Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza 50018, Spain.; Institute for Health Research Aragón (IIS Aragón), Zaragoza 50009, Spain.; CIBER-BBN, ISCIII, Zaragoza 50018, Spain.

DOI: 10.1021/acsbiomaterials.3c01302

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MeSH Terms: Humans, Constriction, Cells, Cultured, Myocytes, Cardiac, Myocardial Ischemia, Ischemia