The effect of 2D culture on stemness-related transcriptome in shed microvesicles of mesenchymal stem cells

Abstract

Abstract Background Over the past years, the applications of mesenchymal stem cells (MSCs) grow rapidly in medicine as a promising option for regenerative medicine. So, increasing demands for MSCs and their derivations as cell-based drugs in the market are anticipated. One of the major challenges in MSC therapy is possible alterations in the stemness potential of MSCs during in vitro culture and proliferation. Microvesicles (MVs) reflect the transcriptional status of cells of origin. This study aimed to investigate the impact of 2D culture on stemness characteristics of MSCs by focusing on transcriptome in shed MVs.Methods and Results MVs were isolated from rat bone marrow MSCs using ultracentrifugation. Transmission electron microscopy and dynamic light scattering were applied to characterizing MVs. Then, some markers related to stemness and differentiation were evaluated in shed MVs during 10-day culture. Meanwhile the cell proliferation and apoptosis were assessed in cultured MSCs. Based on our results, not only MSCs-derived MVs preserved somewhat their stemness contents but also MSCs maintained proliferative during 10-day 2D culture.Conclusion Taken together, 2D culture had no significant effect on the biological state of MSCs which strengthened their applications in cell therapies. Further studies are needed to identify the protein component of MVs to give some benefits for biomimetic approaches.