中国寄生虫学与寄生虫病杂志 ›› 1994, Vol. 12 ›› Issue (2): 81-84.

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间日疟原虫红细胞外期体外培养的研究

罗树宏,刘多,叶炳辉,舒衡平,付冉定   

  1. 湖南医科大学; 南京医科大学
  • 出版日期:1994-05-31 发布日期:2017-01-06
  • 基金资助:
    国家自然科学基金

IN VITRO CULTIVATION OF THE EXOERYTHROCYTIC STAGE OF PLASMODIUM VIVAX (SOUTHERN CHINA ISOLATE)

  • Online:1994-05-31 Published:2017-01-06

摘要: 用人肝癌细胞在体外培养间日疟原虫红细胞外期获得成功,在国内首次建立肝癌细胞—间日疟原虫红细胞外期体外培养系统。在无菌条件下解剖受感染的斯氏按蚊唾腺,制成子孢子悬液,并接种到肝癌细胞中进行体外培养,7d后在培养物中可见红细胞外期裂殖体。第10d,在培养物中加入正常人O型红细胞,继续培养,14d后分离红细胞制成涂片,可见大量环状体和早期大滋养体。提示此种肝癌细胞可作为我国间日疟原虫红细胞外期体外培养的宿主细胞。

关键词: 间日疟原虫, 红细胞外期, 体外培养

Abstract: An in vitro culture system for the exoerythrocytic(EE)stage of Plasmodium vivax was first developed in our laboratory in China. Anopheles stephensi mosquitoes were infected by membrane feeding with heparinized blood from a volunteer.After 14 18 days,the mosquito salivary glands were aseptically dissected in culture medium and ground in a tissue grinder to form sporozoite suspension.Sporozoites were counted and added to the cultures of monolayer hepatoma cells at the number of 4.9 ×10 4-1 3×10 5 per cover glass.Sometimes,8 10 pairs of infected gland were added directly to the cultured cells.On day 7,EE schizonts of P.vivax were found in cultures.In addition,normal erythrocytes (type o)were added to the cultures on day 10 at a concentration of 10 8 per dish.Fourteen days later,erythrocytes in culture supernatant were collected and thin blood films were made.Numerous intra erythrocytic P.vivax parasites were identified on the films after Giemsa staining. Most intra erythrocytic forms were rings and early large trophozoites.Schuffers dots were present in many of the infected cells which were pale and obviously enlarged.These results indicated that the in vitro hepatic cycle of P.vivax was establisthed.

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