中国组织工程研究2017，Vol.21Issue(33)：5375-5381,7.DOI:10.3969/j.issn.2095-4344.2017.33.020
低氧微环境促进人嗅黏膜间充质干细胞的增殖及其机制
Hypoxic microenvironment promotes the proliferation of human olfactory mucosa mesenchymal stem cells and its associated mechanism
摘要
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.关键词
干细胞/培养/人嗅黏膜间充质干细胞/低氧/低氧诱导因子/增殖/国家自然科学基金分类
医药卫生引用本文复制引用
卓毅,李璇,段答,葛丽特,袁挺,吴沛,王昊,龙浪,卢明..低氧微环境促进人嗅黏膜间充质干细胞的增殖及其机制[J].中国组织工程研究,2017,21(33):5375-5381,7.基金项目
国家自然科学基金(81371358) （81371358）
湖南省自然科学基金(14JJ2060) （14JJ2060）
湖南省教育厅研究生创新项目(CX2015B188) the National Natural Science Foundation of China, No. 81371358 （CX2015B188）
the Natural Science Foundation of Hunan Province, No. 14JJ2060 （）
the Postgraduate Innovative Project of Hunan Provincial Education Department, No. CX2015B188 （）
