Abstract
Cell therapy improves cardiac function. Few cells have been investigated more extensively or consistently shown to be more effective than c-kit sorted cells; however, c-kit expression is easily lost during passage. Here, our primary goal was to develop an improved method to isolate c-kitpos cells and maintain c-kit expression after passaging. Cardiac mesenchymal cells (CMCs) from wild-type mice were selected by polystyrene adherence properties. CMCs adhering within the first hours are referred to as rapidly adherent (RA); CMCs adhering subsequently are dubbed slowly adherent (SA). Both RA and SA CMCs were c-kit sorted. SA CMCs maintained significantly higher c-kit expression than RA cells; SA CMCs also had higher expression endothelial markers. We subsequently tested the relative efficacy of SA vs. RA CMCs in the setting of post-infarct adoptive transfer. Two days after coronary occlusion, vehicle, RA CMCs, or SA CMCs were delivered percutaneously with echocardiographic guidance. SA CMCs, but not RA CMCs, significantly improved cardiac function compared to vehicle treatment. Although the mechanism remains to be elucidated, the more pronounced endothelial phenotype of the SA CMCs coupled with the finding of increased vascular density suggest a potential pro-vasculogenic action. This new method of isolating CMCs better preserves c-kit expression during passage. SA CMCs, but not RA CMCs, were effective in reducing cardiac dysfunction. Although c-kit expression was maintained, it is unclear whether maintenance of c-kit expression per se was responsible for improved function, or whether the differential adherence property itself confers a reparative phenotype independently of c-kit.
Highlights
Following myocardial infarction, the loss of contractile units causes left ventricular dysfunction and is the major barrier to treating heart failure
Many groups have studied the role of c-kit-sorted cardiac cells for treatment of heart failure, data regarding the maintenance of c-kit expression during expansion is rare
The primary goal of this study was to establish a reproducible protocol to maintain c-kit expression during passage of Cardiac mesenchymal cells (CMCs), and to confirm whether the resulting cells had reparative potential
Summary
The loss of contractile units causes left ventricular dysfunction and is the major barrier to treating heart failure. C-Kit Stabilization in CMCs evidence supports the efficacy of cell therapy in preclinical models (Bolli and Ghafghazi, 2015; Golpanian et al, 2016), and the results of clinical trials have been encouraging, suggesting that cell therapy is both safe and effective in patients with heart failure (Khan et al, 2016). Adoptive transfer of various types of progenitor/mesenchy mal/stromal/stem cells attenuates cardiac dysfunction in preclinical heart failure models (Keith and Bolli, 2015). The receptor tyrosine kinase, c-kit, is a putative marker of cardiac stem/progenitor cells, and some studies have shown that c-kit sorted cells transdifferentiate to cardiomyocytes (Nadal-Ginard et al, 2014; Torella et al, 2014; Waring et al, 2014), others have been unable to reproduce this finding (Keith and Bolli, 2015). If c-kit expression is necessary for the effectiveness of c-kit cells, we should be intentional about identifying a reproducible approach to stabilize c-kit expression during passage—this was a primary goal of the present study
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