论文部分内容阅读
目的分离并诱导培养胎鼠皮肤来源肥大细胞。方法分离14~16 d胎鼠皮肤细胞并加入重组小鼠白细胞介素3(IL-3)和重组小鼠干细胞因子共同诱导其中的肥大细胞分化成熟。细胞培养14 d后,经密度梯度离心法得到成熟肥大细胞。采用甲苯胺蓝染色或类胰蛋白酶免疫荧光细胞化学染色检测判断肥大细胞纯度,流式细胞术检测Ig E的高亲和力受体(FcεRⅠ)和CD117进一步鉴定肥大细胞纯度。结果培养14 d并纯化后,细胞经甲苯胺蓝及免疫荧光染色可见胞质内大量颗粒,FcεR I+CD117+细胞达97%。结论成功从胎鼠皮肤分离并获得大量高纯度成熟的肥大细胞。
Objective To isolate and induce fetal skin derived mast cells. Methods Fetal rat skin cells were isolated from 14 to 16 days and recombinant mouse interleukin 3 (IL-3) and recombinant mouse stem cell factor were added to induce mast cell differentiation and maturation. After 14 days of cell culture, mature mast cells were obtained by density gradient centrifugation. The purity of mast cells was determined by toluidine blue staining or tryptase immunofluorescence cytochemical staining. The purity of mast cells was further identified by flow cytometry analysis of Igε high affinity receptor (FcεRI) and CD117. Results After cultured for 14 days and purified, the cells were stained with toluidine blue and immunofluorescence to observe a large number of granules in the cytoplasm. The number of FcεR I + CD117 + cells reached 97%. Conclusion A large number of high-purity mature mast cells were successfully isolated from fetal rat skin.