Hypoxic microenvironment promotes the proliferation of human olfactory mucosa mesenchymal stem cells and its associated mechanism
10.3969/j.issn.2095-4344.2017.33.020
- VernacularTitle:低氧微环境促进人嗅黏膜间充质干细胞的增殖及其机制
- Author:
Yi ZHUO
1
;
Xuan LI
;
Da DUAN
;
te Li GE
;
Ting YUAN
;
Pei WU
;
Hao WANG
;
Lang LONG
;
Ming LU
Author Information
1. 湖南师范大学第二附属医院(解放军第163医院)神经外科
- From:
Chinese Journal of Tissue Engineering Research
2017;21(33):5375-5381
- CountryChina
- Language:Chinese
-
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.
