Sequestration of Plasmodium falciparum-infected erythrocytes (IEs) is responsible for the pathophysiology of placental malaria, leading to serious complications such as intrauterine growth restriction and low birth weight. However, it is an experimental challenge to study the biology of human placenta. Conventional cell culture-based in vitro placental models rely on immunostaining techniques and high-magnification microscopy is limited in providing real-time quantitative analysis. Impedimetric sensing in combination with cell culture may offer a useful tool. In this paper, we report that real-time label-free measurement of cellular electrical impedance using xCELLigence technology can be used to quantify the proliferation, syncytial fusion, and long-term response of BeWo cells to IEs cytoadhesion. Specifically, we optimized key experimental parameters of cell seeding density and concentration of forskolin, a compound used to promote cell syncitiation, based on electrical signals and immunostaining results. Prolonged time of infection with IEs that led to cell-cell junction vanishment in BeWo cells and release of inflammatory cytokines were monitored in real time by continuous change in electrical impedance. The results suggest that the impedimetric technique is sensitive and can offer new opportunities for the study of cellular responses of trophoblast cells to IEs. The developed system can provide potentially a high-throughput screening tool of anti-adhesion or anti-inflammatory drugs for placental malaria infections.

中文翻译：

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实时滋养细胞检测恶性疟原虫疟疾中滋养层-红细胞相互作用的体外滋养层检测方法的优化。

螯合恶性疟原虫感染的红细胞（IEs）导致胎盘疟疾的病理生理，导致严重的并发症，如宫内生长受限和低出生体重。然而，研究人类胎盘生物学是一项实验性挑战。基于常规细胞培养的体外胎盘模型依赖于免疫染色技术，高放大倍数显微镜在提供实时定量分析方面受到限制。阻抗传感与细胞培养相结合可能会提供有用的工具。在本文中，我们报道了使用xCELLigence技术对细胞电阻抗进行实时无标签测量可用于量化BeWo细胞对IEs细胞粘附的增殖，合胞融合和长期反应。特别，我们根据电信号和免疫染色结果优化了细胞接种密度和毛喉素浓度（用于促进细胞趋同的化合物）的关键实验参数。通过持续不断的电阻抗变化实时监测IEs的延长感染时间，这些IEs导致BeWo细胞中的细胞间连接消失以及炎性细胞因子的释放。结果表明，阻滞技术是敏感的，并可以为滋养层细胞对IEs的细胞反应研究提供新的机会。开发的系统可以为胎盘疟疾感染提供潜在的高通量抗粘连或抗炎药物筛选工具。基于电信号和免疫染色结果。通过持续不断的电阻抗变化实时监测IEs的延长感染时间，这些IEs导致BeWo细胞中的细胞间连接消失以及炎性细胞因子的释放。结果表明，阻滞技术是敏感的，可以为研究滋养层细胞对IEs的细胞反应提供新的机会。开发的系统可以为胎盘疟疾感染提供潜在的高通量抗粘连或抗炎药物筛选工具。基于电信号和免疫染色结果。通过持续不断的电阻抗变化实时监测IEs的延长感染时间，这些IEs导致BeWo细胞中的细胞间连接消失以及炎症性细胞因子的释放。结果表明，阻滞技术是敏感的，可以为研究滋养层细胞对IEs的细胞反应提供新的机会。开发的系统可以为胎盘疟疾感染提供潜在的高通量抗粘连或抗炎药物筛选工具。结果表明，阻滞技术是敏感的，可以为研究滋养层细胞对IEs的细胞反应提供新的机会。开发的系统可以为胎盘疟疾感染提供潜在的高通量抗粘连或抗炎药物筛选工具。结果表明，阻滞技术是敏感的，可以为研究滋养层细胞对IEs的细胞反应提供新的机会。开发的系统可以为胎盘疟疾感染提供潜在的高通量抗粘连或抗炎药物筛选工具。