Paracrine potential of adipose stromal vascular fraction cells to recover hypoxia‐induced loss of cardiomyocyte function

Abstract

Cell-based therapies show promising results in cardiac function recovery mostly through paracrine-mediated processes (as angiogenesis) in chronic ischemia. In this study, we aim to develop a 2D (two-dimensional) in vitro cardiac hypoxia model mimicking severe cardiac ischemia to specifically investigate the prosurvival paracrine effects of adipose tissue-derived stromal vascular fraction (SVF) cell secretome released upon three-dimensional (3D) culture. For the 2D-cardiac hypoxia model, neonatal rat cardiomyocytes (CM) were cultured for 5 days at < 1% (approaching anoxia) oxygen (O ) tension. Typical cardiac differentiation hallmarks and contractile ability were used to assess both the cardiomyocyte loss of functionality upon anoxia exposure and its possible recovery following the 5-day-treatment with SVF-conditioned media (collected following 6-day-perfusion-based culture on collagen scaffolds in either normoxia or approaching anoxia). The culture at < 1% O for 5 days mimicked the reversible condition of hibernating myocardium with still living and poorly contractile CM (reversible state). Only SVF-medium conditioned in normoxia expressing a high level of the prosurvival hepatocyte-growth factor (HGF) and insulin-like growth factor (IGF) allowed the partial recovery of the functionality of damaged CM. The secretome generated by SVF-engineered tissues showed a high paracrine potential to rescue the nonfunctional CM, therefore resulting in a promising patch-based treatment of specific low-perfused areas after myocardial infarction.

Long-Term Severe In Vitro Hypoxia Exposure Enhances the Vascularization Potential of Human Adipose Tissue-Derived Stromal Vascular Fraction Cell Engineered Tissues

“…Previously published results have focused on the paracrine effects of medium conditioned by SVF-patches cultured in severe hypoxia on a two-dimensional (2D) in vitro cardiac model of dysfunctional cardiomyocytes [ 23 ]. Compared to static culture, the secretome released by hypoxia cultured SVF-based patches was indeed enriched with pro-survival factors (such as the hepatocyte growth factor and the insulin growth factor), and therefore, capable of partially restoring the contractility of dysfunctional cardiomyocytes.…”

Construction of engineered myocardial tissues in�vitro with cardiomyocyte‑like cells and a polylactic‑co‑glycolic acid polymer

“…at present, the mechanism of engineered myocardial tissue remains unclear (40)(41)(42). Previous studies have hypothesized that the paracrine effects of the implanted cells are more likely to affect the survival and growth of engineered myocardial tissues (43)(44)(45). However, there are a number of doubts that require additional confirmation.…”

Versatile electrical stimulator for cardiac tissue engineering—Investigation of charge-balanced monophasic and biphasic electrical stimulations

“…With these techniques, the percentage of area positive for Cx-43 and the percentage of area positive for Sarcomeric α-actinin were quantified ( Navaei et al, 2017 ; Lemme et al, 2018 ; Isu et al, 2020 ; Pisanu et al, 2022 ). Finally, to understand the portion of CMs which underwent a good maturation in terms of sarcomeric organization, the percentage of CMs with sarcomeric organization with respect to the total number of CMs was quantified ( Mytsyk et al, 2019 ).…”