日本血吸虫童虫细胞体外培养条件的初步研究  

Preliminary study on the cultural conditions for the cultivation of schistosomula cells of Schistosoma japonicum in vitro

作　　者：龚燕飞[1] 曾庆仁[1] 张祖萍[1] 林雪迟[1] 蔡力汀[1] 言敢威[1] 

机构地区：[1]中南大学湘雅医学院寄生虫学教研室,长沙410013

出　　处：《中国人兽共患病杂志》2005年第2期159-163,124,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金资助项目(30170840)

摘　　要：目的探讨日本血吸虫童虫细胞体外培养条件。方法选取转铁蛋白、核苷酸、碱性成纤维细胞生长因子(bFGF)、黄体酮以及非必需氨基酸与维生素组合等5种促进细胞增殖的因子作为考察因素,每种因素选取添加与不添加两个水平,采用正交设计,以RPMI-1640为基础培养液,配制16种条件培养基对日本血吸虫童虫细胞进行体外培养,通过动态观察和AKP组化染色法及直观法分析法获得不同培养条件影响细胞增殖和细胞活力的结果。结果第2,4,9,13,14孔中的组织出现明显的细胞增殖现象,其细胞AKP检测值也相应较高,各孔中培养细胞的AKP值经直观分析表明促进细胞增殖最佳因素组合为C1B1A1,即转铁蛋白、核苷酸、bFGF。结论转铁蛋白、核苷酸、bFGF对日本血吸虫童虫细胞增殖具有明显促进作用。To explore the excellent cultural conditions for the cultivation of schistosomula cells of Schistosoma japonicum in vitro, 5 growth-promoting factors that promoted the proliferation of cultured cells, including transferrin, nucleotide, basic fibroblast growth factor (b-FGF), progesterone and non-essential amino acid and vitamin combination, were selected as the investigation elements. Two dose levels were chosen for every factor with two levels of addition or without addition. 16 conditional media were based on the RPM1-1640 used as the basal culture medium, and compounded according to orthogonal design. The schistosomula cells of S.japonicum were cultivated in these 16 culture media, the growth and proliferation of the cultured cells were determined by dynamic observations and the cell viability was assayed by AKP staining, in which the results of AKP assessment were intuitionally analyzed.The experimental results showed that the cultured tissue cells in cultivation plate wells No.2,4,9,13,and 14 appeared to be obviously proliferated and possessed higher AKP values than other cells. By analyzing the AKP values among all the cultured cells, it was demonstrated that the best factor combination to promote the proliferation of cultured cells was C1B1A1,which was transferrin, nucleotide and b-FGF combination. It concludes that the transferrin,nucleotide and b-FGF combination can promote obviously the proliferation of cultured cells from schistsomula cells of S.japonicum.

关 键 词：日本血吸虫童虫 细胞增殖 B-FGF 细胞体外培养 转铁蛋白 AKP 核苷酸 种条 必需氨基酸 培养液 

分 类 号：R383.24[医药卫生—医学寄生虫学] S831[医药卫生—基础医学]