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目的探讨细胞固定对鉴别支气管上皮细胞和嗜酸粒细胞两种细胞共同培养过程中细胞代谢产物来源的作用及其机制。方法用1%多聚甲醛固定嗜酸粒细胞和支气管上皮细胞系BEAS-2B细胞;通过锥虫蓝、伊红细胞染色,观察多聚甲醛固定对细胞形态、结构、活力和与正常细胞接触培养过程中与正常细胞黏附能力的影响;用酶联免疫吸附测定(ELISA)方法定量分析固定细胞与正常细胞共同培养过程中对白细胞介素(IL)-6释放的影响。结果多聚甲醛固定后支气管上皮细胞和嗜酸粒细胞均变为死亡细胞,但其形态、结构及嗜酸粒细胞中颗粒着色无明显变化;多聚甲醛固定的BEAS-2B细胞与正常及固定的嗜酸粒细胞接触培养仅有极少量细胞发生黏附,而多聚甲醛固定的嗜酸粒细胞与正常BEAS-2B细胞接触培养则有较大数量的细胞发生黏附,但黏附细胞数量少于两种正常细胞的共同培养;固定后的嗜酸粒细胞与正常BEAS-2B细胞共同培养过程中可诱导BEAS-2B细胞释放IL-6。但与嗜酸粒细胞单独培养比较,固定后BEAS-2B细胞与正常嗜酸粒细胞共同培养对细胞培养上清液中IL-6浓度无明显影响。结论经多聚甲醛固定后,细胞死亡并丧失其代谢功能,但其表面仍保留可诱导其他细胞活化的结构成分,此技术可用于两种或多种细胞共同培养过程中代谢产物来源的鉴别。
OBJECTIVE: To investigate the role of cell fixation in differentiating the sources of cellular metabolites in the co-culture of bronchial epithelial cells and eosinophils. Methods The eosinophil and bronchial epithelial cell line BEAS-2B cells were fixed with 1% paraformaldehyde. The cells were stained with trypan blue and eosin, and the cell morphology, structure, vitality and contact culture with normal cells were observed (ELISA) was used to quantitatively analyze the influence of interleukin (IL) -6 release during co-culture of fixed cells and normal cells. Results After paraformaldehyde fixation, both bronchial epithelial cells and eosinophils became dead cells, but their morphology, structure and eosinophilic granule staining had no significant changes. Paraformaldehyde fixed BEAS-2B cells were normal and fixed Of eosinophils contact cultured only a very small amount of cell adhesion, and paraformaldehyde-fixed eosinophils and normal BEAS-2B cells in contact culture a greater number of cells adhesion, but the number of adherent cells less than two Normal cells were co-cultured. Immobilized eosinophils co-cultured with normal BEAS-2B cells induced the release of IL-6 from BEAS-2B cells. However, compared with eosinophil culture alone, the co-culture of BEAS-2B cells with normal eosinophils did not significantly affect the concentration of IL-6 in the cell culture supernatant. Conclusions After paraformaldehyde fixation, the cells die and lose their metabolic function, but the surface still retains the structural components that can induce the activation of other cells. This technique can be used to identify the sources of metabolites in the co-culture of two or more cells.