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目的:本研究旨在运用一种改良的甲苯胺蓝染色方法鉴定体外分离培养的肥大细胞形态,为肥大细胞研究提供简便的检测方法。方法:体外诱导分化培养小鼠骨髓来源的肥大细胞,4周后收集细胞、固定、染色。比较不同固定温度及时间对甲苯胺蓝染色效果的影响,确定最佳条件。结果:SCF和IL-3体外培养诱导出小鼠骨髓来源肥大细胞。肥大细胞用改良的甲苯胺蓝染色后细胞着色效果较好,细胞多呈圆形或椭圆形且胞膜较完整,胞浆中充满大量的紫红色颗粒。结论:本研究成功运用一种适用于体外培养小鼠骨髓源肥大细胞的甲苯胺蓝染色法,并发现肥大细胞在37℃充分固定后进行染色,可以降低肥大细胞发生脱颗粒。此操作方法稳定性好、简便,适用于体外培养的肥大细胞形态学观察。
OBJECTIVE: This study aimed to identify the morphology of mast cells isolated and cultured in vitro by using an improved toluidine blue staining method, providing a simple and convenient method for the study of mast cells. Methods: Mouse bone marrow-derived mast cells were induced in vitro and collected after 4 weeks. The cells were fixed and stained. The effects of different fixed temperature and time on the toluidine blue staining were compared to determine the best conditions. Results: Mouse bone marrow-derived mast cells were induced by SCF and IL-3 in vitro. Mast cells with modified toluidine blue staining cells better staining, cells were mostly round or oval and more complete cell membrane, cytoplasm filled with a large number of purple particles. CONCLUSION: Toluidine blue staining of mouse bone marrow-derived mast cells was successfully used in this study. Mast cells were stained well after fixation at 37 ℃, which could reduce the degranulation of mast cells. This method is stable, simple, suitable for in vitro culture of the morphology of mast cells.