肥大细胞系LAD2的培养及肥人细胞脱颗粒模型的建立

The Cultivation of Mast Cell Line LAD2 and Establishment of Mast Cell Degranulation Model

目的培养人肥大细胞系LAD2和建立IgE介导的肥大细胞脱颗粒模型,为研究Ⅰ型超敏反应提供体外实验模型。方法噻唑蓝(MTT)检测细胞生长曲线,细胞染色及电镜观察,Biotin-IgE致敏LAD2细胞,链霉亲和素(Streptavidin)攻击,测定组胺及β-氨基己糖苷酶的释放,判断LAD2细胞的激活效应。结果 MTT检测LAD2细胞的倍增时间为10d;细胞染色及电镜发现细胞内有致密颗粒,随着BiotinIgE致敏浓度的升高,组胺及β-氨基己糖苷酶释放量均逐渐升高。结论人肥大细胞系LAD2培养成功,建立了由IgE介导的脱颗粒模型,为进一步研究Ⅰ型超敏反应奠定基础。

Objective To cultivate human mast cell line LAD2 and establish the model of mast cell degranulation mediated by IgE, providing an experimental model for exploring Type I Hypersensitivity in vitro. Methods The growth activity of LAD2 cells was detected by MTr method. LAD2 cells were observed by staining and elec- tron microscope. LAD2 cells were sensitized by biotin-IgE, activated by streptavidin. Effects of cell activation were evaluated by measuring histamine and β-Hexosaminidase release. Results LAD2 cell doubling time was 10 days as determined by MT-F; Cell staining and electron microscope demonstrated that cells had dense granules in cytoplasm; The releases of histamine and β-Hexosaminidase were increased as the concentration of Biotin-IgE was increased. Conclusion LAD2 cell lines were cultivated and passaged successfully. The model of mast cell degranulation was established, which could be used for investigation of Type I Hypersen- sitivity in vitro.