山东医药2018，Vol.58Issue(20)：5-8,4.DOI:10.3969/j.issn.1002-266X.2018.20.002
骨髓间充质干细胞条件培养基对小鼠肺泡上皮细胞增殖的影响及机制
Effects of bone marrow mesenchymal stem cells conditioned medium on proliferation of mouse alveolar epithelial cells and its mechanism
摘要
Abstract
Objective To investigate the effects of bone marrow mesenchymal stem cells conditioned medium ( BM-SCs-CM) on the proliferation of mouse alveolar epithelial cells and its mechanism .Methods Mouse bone marrow mesen-chymal stem cells (BMSCs) were isolated and cultured in vitro, and the cell surface markers were identified by flow cytom-etry.The results showed that the positive rate of CD 44 expression was 92.5％, and the negative expression rate of CD 34 was 97.3％, suggesting the isolated and cultured cells were BMSCs with high purity .When the second generation cell fu-sion reached 70％to 80％, BMSCs-CM was extracted .Human alveolar epithelial cells A 549 were randomly divided into four groups.The control group was cultured in DMEM/F12 medium containing 10％serum, the serum-free group was cul-tured in serum-free DMEM/F12 medium, the BMSCs group was co-cultured with serum-free DMEM/F12 medium in Tran-swells that had been inoculated with mouse BMSCs , and the BMSCs-CM group was cultured with acquired mouse BMSCs-CM.After 24 hours of culture, the relative viability of the cells in each group was measured by using the CCK-8.The in vitro cultured alveolar type Ⅱcells ATⅡwere randomly divided into the control group and BMSCs-CM group.The cells in the control group were cultured in serum-free DMEM/F12 medium for 24 h, and the cells in the BMSCs-CM group were cultured for 24 h in mouse BMSCs-CM.RT-PCR was used to detect the relative expression of sodium channel protein α(α-ENaC).Results The relative survival rates of the serum-free group, the BMSCs group, and the BMSCs-CM group were (80.70 ±0.70)％, (85.83 ±1.98)％, and (91.28 ±1.91)％, respectively.Both the BMSCs group and the BMSCs-CM group were higher than the serum-free group (P<0.05 or P<0.01).The relative expression of α-EnaC in the control group and the BMSCs-CM group was 1.00 ±0.00 and 3.16 ±1.50, respectively;significant difference was found between the two groups (P<0.05).Conclusion BMSCs-CM can promote the proliferation of alveolar epithelial cells by promoting the expression of α-ENaC in alveolar type Ⅱcells.关键词
肺泡上皮细胞/肺泡Ⅱ型细胞/骨髓间充质干细胞条件培养基/钠通道/钠通道蛋白α/小鼠Key words
alveolar epithelial cells/alveolar type Ⅱcells/bone marrow mesenchymal stem cells conditioned medi-um/sodium channel/sodium channel protein α/mouse分类
医药卫生引用本文复制引用
周祉妤,昌建鈞,候亚鹏,丁炎,聂宏光..骨髓间充质干细胞条件培养基对小鼠肺泡上皮细胞增殖的影响及机制[J].山东医药,2018,58(20):5-8,4.基金项目
国家自然科学基金资助项目(81270098,81670010) （81270098,81670010）
辽宁省高等学校基本科研项目(LQNK201745). （LQNK201745）
