The development of sexual stage malaria gametocytes in a Wave Bioreactor.
The development of sexual stage malaria gametocytes in a Wave Bioreactor.
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DOI：
10.1186/s13071-017-2155-z
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发表时间：
2017-05-02
影响因子：
3.2
通讯作者：
Dalton JP
中科院分区：
文献类型：
--
作者：
Demanga CG;Eng JWL;Gardiner DL;Roth A;Butterworth A;Adams JH;Trenholme KR;Dalton JP
阻止疟疾配子体在红细胞中的发育或在蚊子肠道中受精，可以防止蚊媒的感染和疾病向人类宿主的传播。“传播阻断”战略是未来疟疾控制的一个组成部分。然而，缺乏强大的培养系统来大量生产恶性疟原虫配子体，限制了我们对有性期疟疾生物学的了解，并使疫苗或化疗药物的发现变得更加困难。利用Wave Bioreector TM 20/50 EHT培养系统，建立了一种简便、低维护的诱导恶性疟原虫向配子体发生承诺的方法。通过对培养条件的优化，可在2周内获得成熟的V期配子体。我们报道了一种在Wave生物反应器中用N-乙酰氨基葡萄糖(10 MM)诱导配子体发生的简单方法。通过保持培养14-16天，在1个月的L培养中产生了多达1亿个配子体(阶段V)。用磁激活细胞分选柱(MACS)分离的配子细胞等份冷冻保存。这些蚊子通过解冻而重新焕发活力，并被证明保留了它们排泄和感染蚊子(斯氏按蚊)的能力。在符合GMP的条件下，在Wave生物反应器中生产配子体，不仅有助于配子体的细胞、发育和分子研究，而且可以高通量筛选新的抗疟疾药物，并可能开发全细胞配子体或基于子孢子的疫苗。本文的在线版本(doi：10.1186/s13071-017-2155-z)包含补充材料，授权用户可以使用。
Blocking malaria gametocyte development in RBCs or their fertilization in the mosquito gut can prevent infection of the mosquito vector and passage of disease to the human host. A ‘transmission blocking’ strategy is a component of future malaria control. However, the lack of robust culture systems for producing large amounts of Plasmodium falciparum gametocytes has limited our understanding of sexual-stage malaria biology and made vaccine or chemotherapeutic discoveries more difficult. The Wave BioreactorTM 20/50 EHT culture system was used to develop a convenient and low-maintenance protocol for inducing commitment of P. falciparum parasites to gametocytogenesis. Culture conditions were optimised to obtain mature stage V gametocytes within 2 weeks in a large-scale culture of up to a 1 l. We report a simple method for the induction of gametocytogenesis with N-acetylglucosamine (10 mM) within a Wave Bioreactor. By maintaining the culture for 14–16 days as many as 100 million gametocytes (stage V) were produced in a 1 l culture. Gametocytes isolated using magnetic activated cell sorting (MACS) columns were frozen in aliquots for storage. These were revitalised by thawing and shown to retain their ability to exflagellate and infect mosquitoes (Anopheles stephansi). The production of gametocytes in the Wave Bioreactor under GMP-compliant conditions will not only facilitate cellular, developmental and molecular studies of gametocytes, but also the high-throughput screening for new anti-malarial drugs and, possibly, the development of whole-cell gametocyte or sporozoite-based vaccines. The online version of this article (doi:10.1186/s13071-017-2155-z) contains supplementary material, which is available to authorized users.