A microfluidics assay to study invasion of human placental trophoblast cells.
A microfluidics assay to study invasion of human placental trophoblast cells.
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DOI：
10.1098/rsif.2017.0131
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发表时间：
2017-05
期刊：
影响因子：
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通讯作者：
Oyen ML
中科院分区：
文献类型：
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作者：
Abbas Y;Oefner CM;Polacheck WJ;Gardner L;Farrell L;Sharkey A;Kamm R;Moffett A;Oyen ML
先兆子痫、胎儿生长受限和死产是全世界主要的妊娠疾病。这些疾病的根本发病机制是胎盘发育缺陷，其特征是绒毛外胎盘滋养层细胞侵入子宫动脉的能力不足。如何控制滋养层入侵仍然是一个悬而未决的问题，但它受到母体子宫免疫细胞（称为蜕膜自然杀伤细胞）的影响。在这里，我们描述了一种体外微流体侵袭试验来研究原代人滋养层细胞的迁移。尽管分离初级滋养层细胞存在挑战，但每个实验都可以使用少量细胞进行，从而可以对人类样本进行研究。将细胞暴露于化学梯度中，并使用实时高分辨率成像在三维微环境中进行跟踪，从而获得细胞迁移的动态读数，例如方向性、运动性和速度。使用分离的滋养层和粒细胞-巨噬细胞集落刺激因子（一种由激活的蜕膜自然杀伤细胞产生的细胞因子）的梯度来验证微流体系统。与之前的检测相比，该微流体模型提供了滋养层迁移动力学的详细分析，并且可以在未来进行修改，以在体外研究人类滋养层在胎盘形成过程中的行为。
Pre-eclampsia, fetal growth restriction and stillbirth are major pregnancy disorders throughout the world. The underlying pathogenesis of these diseases is defective placentation characterized by inadequate invasion of extravillous placental trophoblast cells into the uterine arteries. How trophoblast invasion is controlled remains an unanswered question but is influenced by maternal uterine immune cells called decidual natural killer cells. Here, we describe an in vitro microfluidic invasion assay to study the migration of primary human trophoblast cells. Each experiment can be performed with a small number of cells making it possible to conduct research on human samples despite the challenges of isolating primary trophoblast cells. Cells are exposed to a chemical gradient and tracked in a three-dimensional microenvironment using real-time high-resolution imaging, so that dynamic readouts on cell migration such as directionality, motility and velocity are obtained. The microfluidic system was validated using isolated trophoblast and a gradient of granulocyte-macrophage colony-stimulating factor, a cytokine produced by activated decidual natural killer cells. This microfluidic model provides detailed analysis of the dynamics of trophoblast migration compared to previous assays and can be modified in future to study in vitro how human trophoblast behaves during placentation.