Hypoxic microenvironment promotes the proliferation of human olfactory mucosa mesenchymal stem cells and its associated mechanism

doi:10.3969/j.issn.2095-4344.2017.33.020

Abstract

Abstract:

BACKGROUND: Human olfactory mucosa mesenchymal stem cells (hOM-MSCs) not only have the basic characteristics of mesenchymal stem cells, but also originate from the ectoderm and are prone to differentiate into neurons, which are a kind of ideal seed cells for nerve repair and regeneration. Cells are conventionally cultured in about 21% in vitro, while only 3%-9% oxygen is found in the human body and tissue space. There is still no report on the effect of hypoxia on the proliferation and activity of hOM-MSCs.
OBJECTIVE: To explore whether hypoxic microenvironment can promote hOM-MSCs proliferation and activity and the related mechanism.
METHODS: hOM-MSCs were isolated, cultured and identified by flow cytometry and immunofluorescence. The passage 4 hOM-MSCs were divided into three groups: 21% O₂ group, 3% O₂ group and 3% O₂+20 µmol/L YC-1 (HIF-1α inhibitors) group. Proliferation and apoptosis of hOM-MSCs was detected by flow cytometry after 72 hours of culture. The proliferating cell nuclear antigen protein expression was detected by western blot. The mRNA and protein expression of HIF-1α and TWIST were detected by Q-PCR and western blot.
RESULTS AND CONCLUSION: The purity of hOM-MSCs was up to 97%, as defined by flow cytometry. The proliferation index of 3% O₂ group was higher than the 21% O₂ group (P < 0.05), and cell survival and apoptosis ratio (apoptotic cells included mechanical death + early apoptosis + late apoptosis) between the two groups had no significant difference (P > 0.05). Western blot results showed that the proliferating cell nuclear antigen protein expression in the 3% O₂ group was significantly higher than that in the other groups (P < 0.05). The HIF-1α and TWIST expressions at mRNA and protein levels in the 3% O₂ group were significantly higher than those in the other groups (P < 0.05). To conclude, hypoxic microenvironment can promote the hOM-MSCs proliferation and has no effect on the apoptosis, and the HIF-TWIST signal pathway plays an important role in this progress.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Cell Hypoxia, Olfactory Mucosa, Mesenchymal Stem Cells, Hypoxia-Inducible Factor 1, alpha Subunit, Tissue Engineering

CLC Number:

R394.2

Zhuo Yi, Li Xuan, Duan Da, Ge Li-te, Yuan Ting, Wu Pei, Wang Hao, Long Lang, Lu Ming. Hypoxic microenvironment promotes the proliferation of human olfactory mucosa mesenchymal stem cells and its associated mechanism[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(33): 5375-5381.

Figures/Tables 3

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