3D Human Myocardial Tissue Generation Using Melt Electrospinning Writing of Polycaprolactone Scaffolds and hiPSC-Derived Cardiac Cells

Andrea Sánchez-Bueno; Olalla Iglesias-García; Pilar Montero-Calle; Juan José Gavira; Felipe Prosper; Manuel  M. Mazo

doi:10.3791/67847

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Method Article

3D Human Myocardial Tissue Generation Using Melt Electrospinning Writing of Polycaprolactone Scaffolds and hiPSC-Derived Cardiac Cells

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DOI:

10.3791/67847

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March 28th, 2025

Andrea Sánchez-Bueno¹ , Olalla Iglesias-García¹ , Pilar Montero-Calle¹ , Juan José Gavira² , Felipe Prosper³^,⁴^,⁵ , Manuel M. Mazo¹^,³

¹Biomedical Engineering Program, Enabling Technologies Division, CIMA Universidad de Navarra, and Instituto de Investigación Sanitaria de Navarra (IdiSNA), ²Department of Cardiology, Clínica Universidad de Navarra and Instituto de Investigación Sanitaria de Navarra (IdiSNA), ³Hematology and Cell Therapy Area, Clínica Universidad de Navarra and Instituto de Investigación Sanitaria de Navarra (IdiSNA), ⁴Centro de Investigación Biomédica en Red de Cáncer (CIBERONC) CB16/12/00489, ⁵Hemato-Oncology Program, Cancer Division, CIMA Universidad de Navarra

Summary

A reproducible method is presented to generate 3D myocardial tissues combining melt electrospinning writing (MEW) polycaprolactone (PCL) scaffolds and fibrin hydrogels with hiPSC-derived cardiomyocytes and fibroblasts. This technique offers precise control over scaffold architecture and can be applied in preclinical drug testing and cardiac disease modeling.

Abstract

The development of functional human cardiac tissues holds significant promise for advancing applications in drug screening, disease modeling, and regenerative medicine. This protocol describes the stepwise fabrication of 3D myocardial tissues with advanced mimicry of native cardiac structure by combining melt electrospinning writing (MEW) polycaprolactone (PCL) scaffolds with fibrin hydrogels and human induced pluripotent stem cell (hiPSC)-derived cardiac cells. The process involves embedding a mixture of cardiomyocytes (hiPSC-CMs) and cardiac fibroblasts (hiPSC-CFs) within a fibrin matrix to create mini-tissues, with structural support provided by MEW-generated scaffolds. These fibrillar scaffolds are fabricated at the micro- to nanoscale, allowing for precise control over fiber architecture, which plays a key role in organizing cell distribution and alignment. Meanwhile, the fibrin matrix promotes cell viability and mimics the extracellular environment. Characterization of the generated tissues reveals well-organized sarcomeres within hiPSC-CMs, along with stable contractile activity. The tissues demonstrate consistent spontaneous beating as early as two days post-seeding, with sustained functionality over time. The combination of hiPSC-CFs with hiPSC-CMs enhances the structural integrity of the tissues while supporting long-term cell viability. This approach offers a reproducible, adaptable, and scalable method for creating biomimetic cardiac tissue models, providing a versatile platform for preclinical drug testing, mechanistic studies of cardiac disease, and potential regenerative therapies.

Introduction

Fabrication of functional human cardiac-engineered tissues holds great promise for high-impact applications. These span from the development of modeling drug cardiotoxicity and human cardiac disease to the generation of therapeutic tissues of relevant sizes¹. Although the last 15 years have witnessed significant advances in the field, the fabrication of highly mimetic human myocardium is currently thwarted by difficulties in reproducing the complex structural and mechanical organization of its natural counterpart².

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