RESUMO
To explore whether hypoxic condition could promote the olfactory mucosa mesenchymal stem cells (OM-MSCs) to differentiate into neurons with the olfactory ensheathing cells (OECs) supernatant and the potential mechanisms. Methods: The OM-MSCs and OECs were isolated and cultured, and they were identified by flow cytometry and immunofluorescence. The OM-MSCs were divided into three groups: a 3%O2+ HIF-1α inhibitors (lificiguat: YC-1) + OECs supernatant group (Group A) , a 3%O2 + OECs supernatant group (Group B) and a 21%O2 + OECs supernatant group (Control group). The neurons, which were differentiated from OM-MSCs, were assessed by immunofluorescence test. The mRNA and protein expression of hypoxia-inducible factor-1α (HIF-1α), βIII-tubulin and glial fibrillary acidic portein (GFAP) were detected by quantitative polymerase chain reaction (Q-PCR) and Western blot. The potassium channels were analyzed by patch clamp. Results: The neurons differentiated from OM-MSCs expressed the most amount of βIII-tubulin, and the result of Q-PCR showed that HIF-1α expression in the Group B was significantly higher than that in the other groups (all P<0.05). Western blot result showed that the βIII-tubulin protein expression was significantly higher and GFAP protein expression was obviously decreased in the Group B (both P<0.05). The patch clamp test confirmed that the potassium channels in the neurons were activated. Conclusion: Hypoxic condition can significantly increase the neuronal differentiation of OM-MSCs by the OECs supernatant and decrease the production of neuroglia cells, which is associated with the activation of HIF-1 signal pathway.
Assuntos
Western Blotting , Diferenciação Celular , Fisiologia , Células Cultivadas , Meios de Cultivo Condicionados , Química , Farmacologia , Citometria de Fluxo , Proteína Glial Fibrilar Ácida , Metabolismo , Hipóxia , Subunidade alfa do Fator 1 Induzível por Hipóxia , Metabolismo , Indazóis , Farmacologia , Células-Tronco Mesenquimais , Fisiologia , Neurogênese , Fisiologia , Neuroglia , Metabolismo , Fisiologia , Neurônios , Fisiologia , Mucosa Olfatória , Canais de Potássio , Transdução de Sinais , Tubulina (Proteína) , MetabolismoRESUMO
OBJECTIVE@#To observe the biological characteristics of the human olfactory mucosa mesenchymal stem cells (hOM-MSCs).@*METHODS@#The hOM-MSCs were isolated, cultured and identified in vitro. Scanning electron microscope and transmission electron microscope were used to observe the ultrastructure of hOMMSCs. Th e cells were induced towards adipocyte, osteocyte, neural stem cells, neural-like-cells in vitro.@*RESULTS@#The hOM-MSCs were mainly in spindle shape, arranged with radial colony. The hOMMSCs expressed CD73 and CD90 but no CD34 and CD45. Th e short and thick microvilli processes were seen at the surface of hOM-MSCs by scanning electron microscope, and 2 different cellular morphology of hOM-MSCs were seen under transmission electron microscope. Moreover, the hOMMSCs could be differentiated into adipocyte, osteocyte, neural stem cells and neural cells.@*CONCLUSION@#The hOM-MSCs possess general biological characteristics of MSCs and display multiple differentiation functions. They can be served as ideal seed cells in tissue-engineering for injury repair.