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论文摘要

牙周膜肌成纤维细胞的体外培养及其标志物的表达时效

Culture of human periodontal myofibroblast and time-dependent effect of cell markers

作者:孟耀 刘曼 白丁

Author:Meng Yao, Liu Man, Bai Ding

收稿日期:2014-09-03          年卷(期)页码:2015,42(3):285-289

期刊名称:国际口腔医学杂志

Journal Name:International Journal of Stomatology

关键字:人牙周膜成纤维细胞,肌成纤维细胞,α-平滑肌肌动蛋白,

Key words:human periodontal fibroblast,myofibroblast,α-smooth muscle actin,

基金项目:

国家自然科学基金(30970705)

中文摘要

目的 探讨人牙周膜肌成纤维细胞(MFB)体外培养及其标志物表达的时效性。方法 体外培养人牙周膜成纤维细胞(hPDLF),72 h内以5 μg•L-1终质量浓度的转化生长因子(TGF)-β1诱导hPDLF向MFB转化,免疫细胞化学和免疫组织化学染色检测MFB标志物α-平滑肌肌动蛋白(α-SMA)表达情况。分别进行12、24、48、72、96和120 h的MFB观察,采用流式细胞术观察MFB长时间培养的细胞活性,采用免疫细胞化学观察MFB长时间培养后的α-SMA表达情况。结果 经TGF-β1诱导后α-SMA呈阳性;诱导至120 h,α-SMA仍呈阳性,72 h内其表达稳定。结论 5 μg•L-1终质量浓度的TGF-β1成功诱导人牙周膜细胞向MFB转化。MFB体外培养具有时效性,培养0~72 h,其状态稳定。

英文摘要

Objective This study aimed to investigate the differentiation of human periodontal fibroblasts into myofibroblasts induced by transforming growth factor(TGF)-β1 in vitro and the time-dependent effect of induced myofibroblast(MFB). Methods Human periodontal fibroblasts were cultured with 5 μg•L-1 TGF-β1 for 72 h in vitro to induce to MFB. As an MFB marker, alpha-smooth muscle actin(α-SMA) was examined through immunocytochemistry and immunofluorescence. After induction, MFB was harvested at 12, 24, 48, 72, 96, and 120 h. Cell activity was evaluated through flow cytometry and immunocytochemistry. Results Treatment with 5 μg•L-1 TGF-β1 induced positive α-SMA expression. MFB maintained its phenotype after 120 h, and cell activity was stabilized after 72 h. Conclusion MFB was successfully induced in vitro with 5 μg•L-1 TGF-β1. The cell phenotype was also stabilized within 72 h.

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