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目的:探讨并改进小鼠肠上皮内淋巴细胞分离技术。方法:采用冰浴致肠黏膜上皮脱落的方法,用含DTT的PBS充分振荡后,依次通过80、400目尼龙筛网过滤,观察分析细胞收获率、细胞活率和细胞纯度。结果:每只鼠约20cm小肠可获得(5.6±0.7)×106个上皮内淋巴细胞。上皮内淋巴细胞纯度为(92.21±5.20)%,细胞活率为(90.46±5.71)%。结论:与国内外其他分离方法相比较,该法操作简便,细胞收获率稳定、细胞活率和细胞纯度高,可用于肠上皮内淋巴细胞的相关研究。
Objective: To explore and improve mouse intestinal epithelial lymphocyte separation technology. Methods: The intestinal mucosal epithelial shedding was induced by ice bath. After shaking well in DTT-containing PBS, it was filtered through 80,400 mesh nylon mesh in order to observe the cell harvest rate, cell viability and cell purity. Results: (5.6 ± 0.7) × 106 intraepithelial lymphocytes were obtained in about 20 cm of small intestine per mouse. The purity of intraepithelial lymphocytes was (92.21 ± 5.20)% and the viability of cells was (90.46 ± 5.71)%. Conclusion: Compared with other isolation methods at home and abroad, the method is simple and convenient, the cell harvest rate is stable, the cell viability and cell purity are high, which can be used in the study of intestinal epithelial lymphocytes.