A Pumpless and Tubeless Microfluidic Device Enables Extended In Vitro Development of Cryptosporidium parvum.

IF 3.8 4区医学 Q2 IMMUNOLOGY Open Forum Infectious Diseases Pub Date : 2024-10-16 eCollection Date: 2024-11-01 DOI:10.1093/ofid/ofae625

Samantha Gunasekera, Benjamin Thierry, Edward Cheah, Brendon King, Paul Monis, Jillian M Carr, Abha Chopra, Mark Watson, Mark O'Dea, Una Ryan

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Abstract

Background: The enteric parasite Cryptosporidium remains a treatment challenge for drinking water utilities globally due to its resistance to chlorine disinfection. However, the lack of an in vitro culture system for Cryptosporidium that is both cost-effective and reliable remains a key bottleneck in Cryptosporidium research.

Methods: Here we report that the microfluidic culture of human ileocecal colorectal adenocarcinoma (HCT-8) cells under fluid shear stress enables the extended development of Cryptosporidium parvum. Specifically, the growth of C. parvum in a user-friendly pumpless microfluidic device was assessed using immunofluorescence assays, scanning electron microscopy, and quantitative polymerase chain reaction, which revealed that development continued for 10 days in total.

Results: Oocysts produced within the microfluidic device were infective to fresh HCT-8 monolayers; however, these oocysts were only present at low levels.

Conclusions: We anticipate that such microfluidic approaches will facilitate a wide range of in vitro studies on Cryptosporidium and may have the potential to be further developed as a routine infectivity assessment tool for the water industry.

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无泵、无管微流体设备可扩展副隐孢子虫的体外培养

背景：由于隐孢子虫对氯消毒具有抗药性，因此肠道寄生虫仍然是全球饮用水处理的难题。方法：我们在此报告，在流体剪切应力作用下对人回盲肠结直肠腺癌（HCT-8）细胞进行微流控培养，可使副孢子虫的生长时间延长。具体来说，我们使用免疫荧光测定法、扫描电子显微镜和定量聚合酶链反应评估了无泵微流体装置中副隐孢子虫的生长情况，结果表明副隐孢子虫的生长总共持续了 10 天：结果：微流控装置内产生的卵囊对新鲜的 HCT-8 单层细胞具有感染性，但这些卵囊的数量较少：我们预计这种微流控方法将有助于对隐孢子虫进行广泛的体外研究，并有可能进一步发展成为水行业的常规感染性评估工具。

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来源期刊

Open Forum Infectious Diseases Medicine-Neurology (clinical)

CiteScore

6.70

自引率

4.80%

发文量

630

审稿时长

9 weeks

期刊介绍： Open Forum Infectious Diseases provides a global forum for the publication of clinical, translational, and basic research findings in a fully open access, online journal environment. The journal reflects the broad diversity of the field of infectious diseases, and focuses on the intersection of biomedical science and clinical practice, with a particular emphasis on knowledge that holds the potential to improve patient care in populations around the world. Fully peer-reviewed, OFID supports the international community of infectious diseases experts by providing a venue for articles that further the understanding of all aspects of infectious diseases.