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

[摘要] 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 ofhc-kitpositivecells included immunohistochemical screening with a panel of monoclonal antibodies.Results. Cells, including phase-bright cells identified ashc-kitpositive, spontaneously emigrated from hAA-CHF in suspended explant cultures (SEC) after Day 7. When cocultured with tissue, emigratedhc-kitpositivecells proliferated, first as loosely attached clones and later as multicellular clusters. At Day 21~5% of cells werehc-kitpositive. Between Days 14 and 28hc-kitpositivecells exhibited mesodermal commitment (GATA-4positiveandNKX2.5positive); then after Day 28 cardiac lineages (flk-1positive, smooth muscleactinpositive,troponin-Ipositive, and myosin lightchainpositive).Conclusions.C-kitpositivehCPCs can be derived from atrial tissue of pediatric patients with end-stage HF. SEC is a novel culture method for derivation of migratoryhc-kitpositivecells that favors clinical translation by reducing the need for exogenously added factors to expand hCPCsin vitro.

[学科分类] 生物技术