鸡腹腔巨噬细胞的体外培养与活性分析

Culture and activity analysis of chicken peritoneal macrophages

本试验旨在建立一种适用于鸡腹腔巨噬细胞体外培养的方法,并分析体外培养巨噬细胞的形态特征及活性,以获得数量多、活性高的鸡腹腔巨噬细胞。对5~6周龄伊莎褐蛋公鸡进行腹腔巨噬细胞提取,通过扫描电镜、透射电镜和荧光染色等方法对鸡腹腔巨噬细胞的形态、吞噬活性进行检测。结果显示,成功从5~6周龄伊莎褐蛋公鸡体内分离到腹腔巨噬细胞,数量为(0.62±0.51)×10^6个/羽;细胞提取前24 h向鸡腹腔注射预冷的3%的巯基乙酸盐肉汤能显著增加鸡腹腔巨噬细胞的获得量,数量为(3.54±1.05)×10^6个/羽。鸡腹腔巨噬细胞体外培养36~48 h后呈现出最佳状态,具有完整的形态结构和活跃的吞噬活性。结果表明,3%巯基乙酸盐肉汤诱导能显著提高鸡腹腔巨噬细胞的数量,腹腔巨噬细胞体外培养36~48 h后形态和吞噬活性最佳,可用于后续试验的开展。

This study aimed to establish a suitable method for the chicken peritoneal macrophage culture in vitro,via which to get sufficient number of chicken peritoneal macrophages with high activities.Morphology and activities of peritoneal macrophages isolated from five to six-week-old ISA chickens were detected by scanning electron microscopy,transmission electron microscopy or phagocytosis test.Results show that about(0.62±0.51)×10^6 peritoneal macrophages can be isolated from a five to six-week-old ISA chicken.Intraperitoneal injection of 3%thioglycollate broth to ISA chickens one day in advance significantly increased the number of peritoneal macrophages,which reached(3.54±1.05)×10^6 cells per chicken.Comparatively,the isolated chicken peritoneal macrophages presented the best state after adherent growth for 36 to 48 hours,showing the intact morphological structure and efficient phagocytosis.In conclusion,intraperitoneal injection of 3%thioglycolate broth was an effect way to improve the number of peritoneal macrophages in five to six-week-old ISA chickens.Chicken peritoneal macrophages presented the perfect activities after adherent growth for 36 to 48 hours,which can be used in the next project.