Physiologic expansion of human heart-derived cells enhances therapeutic repair of injured myocardium

Abstract

BackgroundSerum-free xenogen-free defined media and continuous controlled physiological cell culture conditions have been developed for stem cell therapeutics, but the effect of these conditions on the relative potency of the cell product is unknown. As such, we conducted a head-to-head comparison of cell culture conditions on human heart explant-derived cells using established in vitro measures of cell potency and in vivo functional repair.MethodsHeart explant-derived cells cultured from human atrial or ventricular biopsies within a serum-free xenogen-free media and a continuous physiological culture environment were compared to cells cultured under traditional (high serum) cell culture conditions in a standard clean room facility.ResultsTransitioning from traditional high serum cell culture conditions to serum-free xenogen-free conditions had no effect on cell culture yields but provided a smaller, more homogenous, cell product with only minor antigenic changes. Culture within continuous physiologic conditions markedly boosted cell proliferation while increasing the expression of stem cell-related antigens and ability of cells to stimulate angiogenesis. Intramyocardial injection of physiologic cultured cells into immunodeficient mice 1 week after coronary ligation translated into improved cardiac function and reduced scar burden which was attributable to increased production of pro-healing cytokines, extracellular vesicles, and microRNAs.ConclusionsContinuous physiological cell culture increased cell growth, paracrine output, and treatment outcomes to provide the greatest functional benefit after experimental myocardial infarction.

Extracellular Endothelial Cell-Derived Vesicles: Emerging Role in Cardiac and Vascular Remodeling in Heart Failure

“…Interestingly, some subsets of free cell non-coding RNAs, such as mi-RNAs, are normally derived to the target cells by high-density lipoproteins (107), however, the majority of long non-coding RNAs and short chains of mi-RNAs are enriched and stable in EVs and can be delivered by EVs acting as gene regulators (108). Several characteristics of various progenitors cells, which are embedded onto cardiac and vascular remodeling and are expected to carry benefits to the failing heart and vasculature, such as trans-differentiation, paracrine output, migration, survival, are able to be potentially regulated by non-coding RNAs disembarked from endothelial cell-derived EVs (90,109). For instance, endothelial cell-derived EVs through a transfer of long noncoding RNA Mhrt have exhibited the ability to cause acetylation of myocardin, which plays a pivotal role in re-programming cardiac myocytes (50,51).…”

“…Interestingly, there are some mi-RNAs (-146a,−155) that were associated with various metabolic comorbidities (type 2 diabetes mellitus, abdominal obesity, resistance to insulin) among patients with HF and adverse cardiac remodeling (70,108), but the role of endothelial cell-derived EVs in transportation of these molecules still needs to be confirmed further. In contrast, micro-RNA-126 being a component of endothelial cell-derived EVs mediates protein kinase G activity, VCAM-1 expression on the surface of endothelial cells, and increases monocyte recruitment and differentiation (53,90,(109)(110)(111)(112)(113)(114)(115)(116)(117)(118)(119). Several specific mi-RNAs (-92a,−126, and−133) were determined as regulators of microvascular coronary endothelial function and blood coagulation (120,121), and mi-RNA-138 and−155 were negatively associated with NO production and cell-cell communication, respectively (122,123).…”

Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Improves Energetic Status and Cardiomyogenic Differentiation of Human Dilated Myocardium-Derived Primary Mesenchymal Cells

“…It was already agreed that heart tissue is not a post-mitotic organ, constituting cardiomyocytes, fibroblasts (or mesenchymal stromal cells, MSC), and endothelial cells, and having some, albeit limited, regenerative capacity [ 34 , 35 ]. Since the adult human heart has a very limited number of human receptor tyrosine kinase (c-kit) positive stem cells [ 36 ], other types of cells residing in the heart such as human myocardium-derived MSC react to the external stimuli and can be successfully used for cardio regenerative purposes [ 36 , 37 ]. In this study, the HDAC activity and energetic potential of mesenchymal stromal cells (MSC), also named as medicinal signaling cells [ 38 ], derived from human healthy and dilated myocardium biopsies and their response to HDAC class I/II inhibitor SAHA were investigated.…”

Guidance document on Good Cell and Tissue Culture Practice 2.0 (GCCP 2.0)

“…In addition, the height/level of medium above cells determines the accessibility to oxygen and should be recognized as an important factor for cell culture (Al-Ani et al, 2018). The use of closed cell handling chambers to provide constant physiologically relevant oxygen can improve cell function (Mount et al, 2019;Wong et al, 2020).…”