Abstract
The quality and quantity of endothelial progenitor cells (EPCs) are impaired in patients with diabetes mellitus patients, leading to reduced tissue repair during autologous EPC therapy. This study aimed to address the limitations of the previously described serum‐free Quantity and Quality Control Culture System (QQc) using CD34+ cells by investigating the therapeutic potential of a novel mononuclear cell (MNC)‐QQ. MNCs were isolated from 50 mL of peripheral blood of patients with diabetes mellitus and healthy volunteers (n = 13 each) and subjected to QQc for 7 days in serum‐free expansion media with VEGF, Flt‐3 ligand, TPO, IL‐6, and SCF. The vascular regeneration capability of MNC‐QQ cells pre‐ or post‐QQc was evaluated with an EPC colony‐forming assay, FACS, EPC culture, tube formation assay, and quantitative real time PCR. For in vivo assessment, 1 × 104 pre‐ and post‐MNC‐QQc cells from diabetic donors were injected into a murine wound‐healing model using Balb/c nude mice. The percentage of wound closure and angio‐vasculogenesis was then assessed. This study revealed vasculogenic, anti‐inflammatory, and wound‐healing effects of MNC‐QQ therapy in both in vitro and in vivo models. This system addresses the low efficiency and efficacy of the current naïve MNC therapy for wound‐healing in diabetic patients. As this technique requires a simple blood draw, isolation, and peripheral blood MNC suspension culture for only a week, it can be used as a simple and effective outpatient‐based vascular and regenerative therapy for patients with diabetes mellitus.
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