Stem Cell and Regenerative Medicine
Open AccessModulation of Naïve Mesenchymal Stem/Stromal Cells into InsulinProducing Cells by Extracellular Vesicles: Optimization of Coculture Conditions
Authors: Ganna M Metaweh, Abdel-Aziz AF, Sawsan M El-Halawani, Raghda W Magar, Mahmoud M Gabr
Abstract
Aim: This study aimed to optimize the coculture conditions of extracellular vesicles (EVs) derived from surrogate beta cells with naïve mesenchymal stem cells (MSCs).
Methods: MSCs were isolated from liposuction aspirates from three healthy donors. These cells were expanded and differentiated into insulin-producing cells (IPCs). EVs were retrieved from the conditioned media of naïve (uneducated) and differentiated (educated) MSCs by ultracentrifugation. The isolated EVs were characterized by transmission electron microscopy, particle size distribution analysis, and flow cytometry. EVs were cocultured with naïve MSCs under different experimental conditions. Cell preparations were evaluated by immunofluorescence, flow cytometry, and RT-PCR.
Result: the results revealed that 80 µg of educated EVs cocultured with 1x 105 MSCs for 24 h provided the optimal conditions for modulating naïve MSCs into IPCs.
Conclusion: The coculture of EVs derived from IPCs with an appropriate concentration and duration could modulate naïve MSCs into IPCs.
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