NRP2+ human mesenchymal stem cells have stemness-associated properties

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Abstract Background The clinical application of mesenchymal stem cells (MSCs) has garnered attention due to their remarkable capacity to differentiate into adipocytes, chondrocytes, and osteoblasts. However, the quality of MSC culture varies from batch to batch, which poses challenges in ensuring consistent cellular quality across batches. Consequently, it becomes imperative to identify specific markers that can distinguish superior and slightly inferior MSCs. Methods Human bone marrow-derived MSC clones were isolated and subjected to flow cytometry analysis to assess the expression of NRP2, VEGFR, and plexinA1. The osteogenic and adipogenic differentiation potentials were evaluated using Alizarin Red S and Oil Red O staining, respectively. Furthermore, the migration capacity was assessed through the scratch healing assay. Results Nine out of twenty MSC clones significantly expressed NRP2. NRP2-expressing MSC clones (NRP2+ MSCs) retained superior proliferation and differentiation capacities, along with increased migratory capacity compared to non-expressing MSC clones (NRP2− MSCs). In addition, the activation of VEGF-C/NRP2 signaling augmented the potential of MSCs in cell proliferation and differentiation. Conclusion In contrast to NRP2− MSCs, NRP2+ MSCs exhibited superior proliferation, differentiation abilities, and migration capacity. Moreover, the stimulation of VEGF-C/NRP2 signaling further enhanced the proliferation and differentiation rates, indicating a role of NRP2 in the maintenance of MSC stemness. Hence, NRP2 holds potential as a cell surface marker for identifying beneficial MSCs for regenerative medicine.

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