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Background . Human cardiac-derived progenitor cells (hCPCs) have shown promise in treating heart failure (HF) in adults. The purpose of this study was to describe derivation of hCPCs from pediatric patients with end-stage HF.  Methods . At surgery, discarded right atrial tissues (hAA) were obtained from HF patients ( n  = 25; hAA-CHF). Minced tissues were suspended in complete (serum-containing) DMEM. Cells were selected for their tissue migration and expression of stem cell factor receptor (hc-kit). Characterization of hc-kit positive  cells included immunohistochemical screening with a panel of monoclonal antibodies.  Results . Cells, including phase-bright cells identified as hc-kit positive , spontaneously emigrated from hAA-CHF in suspended explant cultures (SEC) after Day 7. When cocultured with tissue, emigrated hc-kit positive  cells proliferated, first as loosely attached clones and later as multicellular clusters. At Day 21 ~ 5% of cells were hc-kit positive . Between Days 14 and 28 hc-kit positive  cells exhibited mesodermal commitment (GATA-4 positive  and NKX2.5 positive ); then after Day 28 cardiac lineages (flk-1 positive , smooth muscle actin positive , troponin-I positive , and myosin light chain positive ).  Conclusions . C-kit positive  hCPCs can be derived from atrial tissue of pediatric patients with end-stage HF. SEC is a novel culture method for derivation of migratory hc-kit positive  cells that favors clinical translation by reducing the need for exogenously added factors to expand hCPCs  in vitro .

Heart Cells with Regenerative Potential from Pediatric Patients with End Stage Heart Failure: A Translatable Method to Enrich and Propagate