Myocardial Tissue Engineering With Cells Derived From Human-Induced Pluripotent Stem Cells and a Native-Like, High-Resolution, 3-Dimensionally Printed Scaffold

Abstract

Rationale Conventional three-dimensional (3D) printing techniques cannot produce structures of the size at which individual cells interact. Objective Here, we used multiphoton-excited, 3-dimensional printing (MPE-3DP) to generate a native-like, extracellular matrix (ECM) scaffold with submicron resolution, and then seeded the scaffold with cardiomyocytes (CMs), smooth-muscle cells (SMCs), and endothelial cells (ECs) that had been differentiated from human induced-pluripotent stem cells (iPSCs) to generate a human, iPSC-derived cardiac muscle patch (hCMP), which was subsequently evaluated in a murine model of myocardial infarction (MI). Methods and Results The scaffold was seeded with ~50,000 human, iPSC-derived CMs, SMCs, and ECs (in a 2:1:1 ratio) to generate the hCMP, which began generating calcium transients and beating synchronously within 1 day of seeding; the speeds of contraction and relaxation and the peak amplitudes of the calcium transients increased significantly over the next 7 days. When tested in mice with surgically induced MI, measurements of cardiac function, infarct size, apoptosis, both vascular and arteriole density, and cell proliferation at week 4 after treatment were significantly better in animals treated with the hCMPs than in animals treated with cell-free scaffolds, and the rate of cell engraftment in hCMP-treated animals was 24.5% at week 1 and 11.2% at week 4. Conclusions Thus, the novel MPE-3DP technique produces ECM-based scaffolds with exceptional resolution and fidelity, and hCMPs fabricated with these scaffolds may significantly improve recovery from ischemic myocardial injury.

3D bioprinted functional and contractile cardiac tissue constructs

“…Compared with the other engineering approaches such as solid substrate seeding [21–23] or hydrogel molding [20,24], 3D bioprinting strategy has inherent advantages in fabricating a highly organized cardiac structure. Although several studies have reported cardiac tissue engineering and bioprinting in the past several years [25–28], the cardiac constructs reported in past were small scale with low-level tissue maturation and disoriented muscle fibers, resulting in weak contractility. Therefore, we utilized 3D bioprinting principle to fabricate uniformly organized, centimeter-scaled cardiac tissues.…”

Stem Cell Therapy in Heart Diseases – Cell Types, Mechanisms and Improvement Strategies

“…Nakatana et al have recently engineered a large tissue construct consisting of cardiomyocytes, endothelial cells and vascular cells which were derived from human iPSCs [453]. Likewise, human ESCs and iPSCs have been utilized for the development of 3-dimesional microtissues formed by SC-derived cardiovascular cells [454,455]. In order to mimick human sized organs, specialized scaffolds are needed that provide characteristics similar to the cardiac ECM, match the strength of native myocardium and allow re-population with cardiac cells.…”

3D bioprinting for cardiovascular regeneration and pharmacology

“…Differentiation protocols have been published for CMs, FBs, endothelial, and epicardial cells. Therefore, iPSCs now offer the opportunity to produce all the essential cellular components of the cardiovascular system in vitro [98]. IPSCs can be directly generated from the reversal conversion (reprogramming) of adult mature cells (FBs, keratinocytes, peripheral blood cells, or renal epithelial cells) via retroviral transduction of stemness transduction factors, such as Oct 3/4, Klf4, Sox2 and c-Myc, as well as using chemicals and reprogramming proteins [99,100].…”

“…Additionally, multiphoton-excited 3D printing was used to generate a native-like extracellular matrix scaffold with submicron resolution, and was able to achieve the individual cell interaction [98]. CMs, smooth muscle cells, and ECs were differentiated from human iPSCs to fabricate the cardiac patch.…”