Addition of Mesenchymal Stem Cells Enhances the Therapeutic Effects of Skeletal Myoblast Cell-Sheet Transplantation in a Rat Ischemic Cardiomyopathy Model

Abstract

Introduction: Functional skeletal myoblasts (SMBs) are transplanted into the heart effectively and safely as cell sheets, which induce functional recovery in myocardial infarction (MI) patients without lethal arrhythmia. However, their therapeutic effect is limited by ischemia. Mesenchymal stem cells (MSCs) have prosurvival/ proliferation and antiapoptotic effects on co-cultured cells in vitro. We hypothesized that adding MSCs to the SMB cell sheets might enhance SMB survival post-transplantation and improve their therapeutic effects. Methods and Results: Cell sheets of primary SMBs of male Lewis rats (r-SMBs), primary MSCs of human female fat tissues (h-MSCs), and their co-cultures were generated using temperature-responsive dishes. The levels of candidate paracrine factors, rat hepatocyte growth factor and vascular endothelial growth factor, in vitro were significantly greater in the h-MSC/r-SMB co-cultures than in those containing r-SMBs only, by real-time PCR and enzyme-linked immunosorbent assay (ELISA). MI was generated by left-coronary artery occlusion in female athymic nude rats. Two weeks later, co-cultured r-SMB or h-MSC cell sheets were implanted or no treatment was performed (n = 10 each). Eight weeks later, systolic and diastolic function parameters were improved in all three treatment groups compared to no treatment, with the greatest improvement in the co-cultured cell sheet transplantation group. Consistent results were found for capillary density, collagen accumulation, myocyte hypertrophy, Akt-signaling, STAT3 signaling, and survival of transplanted cells of rat origin, and were related to poly (ADP-ribose) polymerase-dependent signal transduction. Conclusions: Adding MSCs to SMB cell sheets enhanced the sheets' angiogenesis-related paracrine mechanics and, consequently, functional recovery in a rat MI model, suggesting a possible strategy for clinical applications.

<b>Adipose stem cell sheets improved cardiac function in the rat myocardial infarction, but did not alter cardiac contractile responses to β-adrenergic stim</b><b>ulation </b>

“…The most prominent findings were that ASC sheets improved systolic and diastolic functions of MI hearts under an ex-vivo condition like myoblast cell sheets. Since it has been reported that myoblasts maintain the potential of regenerating skeletal muscle during periods of stress, it suggests that myotubules could directly contribute to improvement of cardiac contraction (19). ASC sheets also improved systolic and diastolic functions of the MI hearts without their differentiation to cardiac myocytes.…”

A Bioengineered Hydrogel System Enables Targeted and Sustained Intramyocardial Delivery of Neuregulin, Activating the Cardiomyocyte Cell Cycle and Enhancing Ventricular Function in a Murine Model of Ischemic Cardiomyopathy

“…These include but are not limited to cell-sheets 30–32 , fibrin patches 33, 34 , alginate scaffolds 35, 36 , and hydrogels 25, 37–39 . The hyaluronic acid hydrogel was utilized in this study because of its previous effectiveness in encapsulating similarly sized peptides and its demonstrated safety in humans.…”

A Tissue-Engineered Chondrocyte Cell Sheet Induces Extracellular Matrix Modification to Enhance Ventricular Biomechanics and Attenuate Myocardial Stiffness in Ischemic Cardiomyopathy

“…LV mass was calculated as a parameter of myocardial hypertrophy. 12,18,19 Wall stress index was defined as a parameter of LV wall stress. 20 All analyses were performed by a single investigator in a group-blinded fashion.…”

“…16,18 The equivalent total protein was loaded onto sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. Antibodies obtained were anti-elastin (AB 21608, 1:200 dilution; Abcam), anti-collagen I (AB34710, 1:1000 dilution; Abcam), anti-collagen II (AB3092, 1:100 dilution; Abcam), anti-matrix metalloproteinase-1 (MMP-1) (AB118529, 1:500 dilution; Abcam), anti-MMP-2 (AB51125, 1:1000 dilution; Abcam), anti-tissue inhibitor of metalloproteinase-1 (TIMP-1) (AF580, 1:500 dilution; R&D), and anti-GAPDH (AB9484; Abcam).…”