Abstract
Recent studies have demonstrated that culture under hypoxia has beneficial effects on mesenchymal stem cells (MSCs). However, there are limitations to achieving a stable condition in conventional hypoxic CO2 incubators. DPSCs are a unique type of MSCs which are promising in many regenerative therapies. In this study, we investigated the ideal hypoxic culture environment for DPSCs using a new system that can provide controlled O2 environment. The effects of hypoxia (3%, 5%) on the stemness properties of DPSCs. Their morphology, proliferation rate, expression of stem cell markers, migration ability, mRNA expression of angiogenic/neurotrophic factors and immunomodulatory genes were evaluated and compared. Additionally, the effect of the discrete secretome on proliferation, migration, and neurogenic induction was assessed. Hypoxic DPSCs were found to be smaller in size and exhibited larger nuclei. 5% O2 significantly increased the proliferation rate, migration ability, expression of stem cell markers (CXCR4 and G-CSFR), and expression of SOX2, VEGF, NGF, and BDNF genes of DPSCs. Moreover, secretome collected from 5%O2 cultures displayed higher stimulatory effects on proliferation and migration of NIH3T3 cells and on neuronal differentiation of SH-SY5Y cells. These results demonstrate that 5%O2 may be ideal for enhancing DPSCs growth, stem cell properties, and secretome trophic effect.
Highlights
Mesenchymal stem cells (MSCs) have been evaluated as a potential tool to treat numerous diseases, including tissue injury, degenerative diseases, and immune disorders
It has been shown recently that secretome collected from adipose-derived MSCs (ADMSCs) cultured under less than 5% O2 contains high levels of granulocyte-macrophage-colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), Interleukin-6 (IL-6), and insulin-like growth factor 1 (IGF-1)[27] and was found to be able to protect myocardial infarct in rat[28]
It was harder to detach Dental pulp stem cells (DPSCs) in hypoxic cultures and took a longer time to obtain single cell suspensions from the detached cells compared to DPSCs in normoxic cultures
Summary
Mesenchymal stem cells (MSCs) have been evaluated as a potential tool to treat numerous diseases, including tissue injury, degenerative diseases, and immune disorders. This is due to their multipotent differentiation capacity[1,2] trophic activity[3,4], immunomodulatory properties[5,6,7] and angiogenic/neurogenic properties[8]. 3% O2 tension in cell culture had positive effects on the in vitro survival and self-renewal of bone marrow stem cells (BMSCs)[19]. Some studies demonstrated that cultivation of DPSCs under hypoxic condition (1–3%) enhances proliferation potential[41] and angiogenic potential[42], and affects the odontoblastic differentiation potential[43]
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