Porous, Ventricular Extracellular Matrix-Derived Foams as a Platform for Cardiac Cell Culture

Abstract

To more closely mimic the native cellular microenvironment, 3D scaffolds derived from the extracellular matrix (ECM) are being developed as alternatives to conventional 2D culture systems. In the present study, we established methods to fabricate nonchemically cross-linked 3D porous foams derived entirely from decellularized porcine left ventricle (DLV) for use as an in vitro cardiac cell culture platform. Furthermore, we explored the effects of physically preprocessing the DLV through mechanical mincing versus cryomilling, as well as varying the ECM concentration on the structure, composition, and physical properties of the foams. Our results indicate that the less highly processed minced foams had a more cohesive and complex network of ECM components, enhanced mechanical properties, and improved stability under simulated culturing conditions. To validate the DLV foams, a proof-of-concept study was conducted to explore the early cardiomyogenic differentiation of pericardial fat adipose-derived stem/stromal cells (pfASCs) on the minced DLV foams relative to purified collagen I gel controls. Differentiation was induced using a modified cardiomyogenic medium (MCM) or through stimulation with 5-azacytidine (5-aza), and cardiomyocyte marker expression was characterized by immunohistochemistry and real-time reverse transcriptase-polymerase chain reaction. Our results indicate that early markers of cardiomyogenic differentiation were significantly enhanced on the DLV foams cultured in MCM, suggesting a synergistic effect of the cardiac ECM-derived scaffolds and the culture medium on the induction of pfASC differentiation. Furthermore, in analyzing the response in the noninduced control groups, the foams were observed to provide a mildly inductive microenvironment for pfASC cardiomyogenesis, supporting the rationale for using tissue-specific ECM as a substrate for cardiac cell culture applications.

Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

“…In the present study, enzymatic digestion with α‐amylase was investigated as an alternative approach to pepsin for generating ECM‐derived coatings. In previous studies, our group has fabricated porous foams and microcarriers from α‐amylase‐digested human DAT, porcine decellularized myocardium, and porcine decellularized dermis . However, to the best of our knowledge, this approach has not yet been applied to synthesize fibrous coatings.…”

Processed Tissue–Derived Extracellular Matrices: Tailored Platforms Empowering Diverse Therapeutic Applications

“…Porous scaffolds can be produced from dissolved polymers, providing interconnected networks that allow for vascularization, cell ingrowth, and nutrient diffusion . While several technologies, including solvent casting/particle leaching and gas foaming, have been utilized to generate porous foams for various applications, the existing reports on dECM‐derived porous scaffold production have used freeze‐drying technology . The process involves dECM pulverization and potentially digestion, followed by homogenization and freeze‐drying.…”

“…For instance, dECM‐derived porous foams can be utilized as a potential platform to study the inductive roles of the ECM in governing cell behavior, including viability, proliferation, and differentiation . The highly porous interconnected architecture leads to increased support of cellular functions, promoting cell adhesion, infiltration, communication, and nutrient diffusion.…”

Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

“…In particular, we have pioneered the use of decellularized adipose tissue (DAT) as a scaffolding platform for adipose regeneration 28 . Moreover, we have established methods for synthesizing 3D microcarriers and porous foams using DAT digested with the proteolytic enzyme pepsin or glycolytic enzyme α-amylase 29 30 31 . Notably, we have demonstrated across all of these scaffold formats that the adipose-derived ECM provides an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem/stromal cells (ASCs) in culture.…”