Chin J Schisto Control ›› 2019, Vol. 31 ›› Issue (3): 294-.

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Observation on growth and reproduction of various Leishmania isolates in different culture media

CHENG Fang-Zhou, YANG Yue-Tao, GAO Chun-Hua, WANG Jun-Yun*   

  1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Key Laboratory of Parasite and Vector Biology, National Health Commission; National Center for International Research on Tropical Diseases, Ministry of Science and Technology; WHO Collaborating Centre for Tropical Diseases, Shanghai 200025, China
  • Online:2019-08-28 Published:2019-08-28

不同利什曼原虫分离株在培养基中生长繁殖观察

程芳洲,杨玥涛,高春花,汪俊云*   

  1. 中国疾病预防控制中心寄生虫病预防控制所、国家卫生健康委员会寄生虫病原与媒介生物学重点实验室、国家级热带病国际联合研究中心、世界卫生组织热带病合作中心(上海 200025)
  • 作者简介:程芳洲,女,硕士研究生。研究方向:病原生物学
  • 基金资助:
    国家重大科技专项(2018ZX10734-404)和国家自然科学基金项目(81472923)

Abstract: Objective To compare the growth and reproduction of the promastigotes of Leishmania isolates from various endemic areas of visceral leishmaniasis in China in various culture media, so as to provide experimental evidence for selecting an appropriate medium for the culture of Leishmania. Methods A total of 3 × 105 promastigotes of KS?2, Cy and JIASHI?5 Leishmania isolates were inoculated into 1 mL NNN medium, 1 mL M199 medium supplemented with 20% fetal bovine serum medium, 1 mL M199 medium supplemented with 20% horse serum medium, and 1 mL brain heart infusion medium containing heme, respectively. All media were placed at 22 ℃ under a sterile condition, and the number of promastigotes was counted continuously for 8 days under a microscope. The growth curve was plotted for the three Leishmania isolates. Results The promastigotes of KS?2, Cy and JIASHI?5 Leishmania isolates all grew and reproduced in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium. The number of promastigotes of KS?2, Cy and JIASHI?5 Leishmania isolates was all significantly higher in the NNN medium than in the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05), and the number of promastigotes of the KS?2 isolate was all significantly greater than that of the Cy and JIASHI?5 isolates in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05). In addition, the promastigotes of KS?2, Cy and JIASHI?5 Leishmania isolates failed to grow and reproduce in the brain heart infusion medium. Conclusions The growth and reproduction of the promastigotes of various Leishmania isolates from various endemic areas of visceral leishmaniasis in China vary in the same culture medium, and the growth and reproduction of a Leishmania isolate vary in different culture media. The NNN medium best fits for the culture of Leishmania isolates in the endemic areas of visceral leishmaniasis in China.

Key words: Leishmania, Visceral Leishmaniasis, Promastigote, Culture medium, In vitro culture

摘要: 目的 比较我国不同类型内脏利什曼病流行区利什曼原虫前鞭毛体在不同培养基中的生长繁殖情况,为选择合适培养基用于利什曼原虫培养提供实验依据。方法 将3 ×105个KS?2、Cy、JIASHI?5株利什曼原虫前鞭毛体分别接种至1 mL NNN培养基、1 mL M199 + 20%胎牛血清培养基、1 mL M199 + 20%马血清培养基及1 mL 脑心浸液培养基(含血红素)中,22 ℃温箱中无菌静置培养,显微镜下连续观察计数8 d,绘制3株利什曼原虫前鞭毛体的生长曲线。 结果 KS?2、Cy、JIASHI?5株利什曼原虫前鞭毛体均能在NNN培养基、M199 + 20%胎牛血清培养基和M199 + 20%马血清培养基中生长繁殖,在NNN培养基中培养不同时间后的3株利什曼原虫前鞭毛体计数均显著高于M199 + 20%胎牛血清培养基和M199 + 20%马血清培养基(P均 < 0.05),在这3种培养基中培养不同时间后的KS?2株利什曼原虫前鞭毛体计数均显著高于Cy和JIASHI?5株(P均 < 0.05)。KS?2、Cy、JIASHI?5株利什曼原虫前鞭毛体均不能在脑心浸液培养基中生长繁殖。结论 分离自我国不同类型内脏利什曼病流行区的利什曼原虫在同一培养基中生长增殖速度有差异,同一利什曼原虫分离株在不同培养基中的生长繁殖速度亦有差异。NNN培养基是最适合我国内脏利什曼病流行区利什曼原虫分离株的培养基。

关键词: 利什曼原虫, 内脏利什曼病, 前鞭毛体, 培养基, 体外培养

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