中国寄生虫学与寄生虫病杂志

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约氏疟原虫红内期体外培养的研究

王兵丽*, 黄家福, 陈凡   

  1. 闽南师范大学生物科学与技术学院, 漳州 363000
  • 出版日期:2017-08-30 发布日期:2017-09-14

In vitro cultivation of ethrythrocytic-stage Plasmodium yoelii

WANG Bing-li*, HUANG Jia-fu, CHEN Fan   

  1. Department of Biology Science and Technology, Minnan Normal University, Zhangzhou, 363000, China
  • Online:2017-08-30 Published:2017-09-14

摘要:

约氏疟原虫(Plasmodium yoelii)红内期的体外连续培养尚未取得理想的效果。本研究在Trager等体外培养恶性疟原虫方法的基础上, 对培养条件进行改进: 将约氏疟原虫放在悬液系统培养仪的小室内, 使用磁性搅拌器搅拌成悬液; 每天更换培养基并向培养基中添加新的网织红细胞; 37 ℃、5% CO2培养箱内常规培养。每日取体外培养物尾静脉注射小鼠, 第3天检查小鼠体内疟原虫感染情况。结果显示, 体外培养的24 h内, 约氏疟原虫的原虫率增长至2.200%, 呈小幅增长, 但随后开始下降。第10天, 未发现被感染的红细胞。第1~9天的体外培养物感染的小鼠均呈阳性, 且第10天的体外培养物感染小鼠呈阴性。第1天体外培养物感染小鼠的原虫率最高, 为54.960%。说明约氏疟原虫红内期的体外培养可以维持9 d, 且体外培养的约氏疟原虫具有感染力。

关键词:  约氏疟原虫, 红内期, 培养, 体外

Abstract:

The persistent in vitro cultivation of ethrythrocytic-stage Plasmodium yoelii has not come to successful results. In this study, we modified the cultivation based on Trager’s method. The modifications included: placing P. yoelii parasites in a small chamber of a suspension system, and agitating them into suspension by a magnetic stirrer; daily replacement of the culture medium and addition of new reticulocytes into the medium; and cultivation at 37 ℃ with 5% CO2. The in vitro cultures were injected daily into mice through tail vein, and the infection status in mice was examined on day 3. Results showed that the proportion of red blood cells with parasite infection increased to 2.200% within 24 h of in vitro cultivation, then decreased till 0% on day 10. As to the mouse model, the mice all showed positive infection from days 1 to 9, with highest rate on day 1 (54.960%), and no infection on day 10. This result suggested that the cultivation of erythrocytic-stage P. yoelii can be maintained up to 9 days in vitro without losing infectivity.
 

Key words: Plasmodium yoelii, Ethrythrocytic stage, Culture, In vitro