Department of Analytical Chemistry , China Pharmaceutical University , Nanjing , Jiangsu 210009 , China.
Department of Biotechnology, Dalian Institute of Chemical Physics , Chinese Academy of Sciences , Dalian , Liaoning 116023 , China.
Anal Chem. 2018 May 1;90(9):5825-5832. doi: 10.1021/acs.analchem.8b00362. Epub 2018 Apr 11.
Despite rapid progresses in single-cell analysis technologies, efforts to control the three-dimensional microenvironment for single cell measurements have been lacking. Here, we report a simple method to incorporate three-dimensional scaffolds, including polyvinylidene fluoride (PVDF) membranes and PVDF membrane replicated analog polydimethylsiloxane, into multiplexed single cell secretomic analysis platforms (including a microwell array and a single cell barcode microchip) to mimic the extracellular physical matrix and mechanical support for single cells. Applying this platform to brain tumor cell line U87 to investigate single cell protein secretion behavior on different substrates, we revealed that single cell protein secretions were regulated differently in three-dimensional (3D) microenvironments. This finding was further verified with intracellular cytokine staining, highlighting the significance of 3D single cell microenvironments. This new single cell biomimetic platform can be easily adaptable to other three-dimensional cell culture scaffolds or other single cell assays and may become a broadly applicable three-dimensional single cell analysis system to study the effect of microenvironment conditions on cellular functional heterogeneity in vitro.
尽管单细胞分析技术取得了快速进展，但对于单细胞测量的三维微环境的控制仍缺乏研究。在这里，我们报告了一种简单的方法，可以将三维支架（包括聚偏二氟乙烯 (PVDF) 膜和 PVDF 膜复制的类似聚二甲基硅氧烷）纳入多重单细胞分泌组学分析平台（包括微井阵列和单细胞条码微芯片）中，以模拟细胞外物理基质和机械支撑对于单细胞。我们将该平台应用于脑肿瘤细胞系 U87，以研究不同基质上的单细胞蛋白分泌行为，结果表明，三维（3D）微环境中，单细胞蛋白分泌受到不同的调控。通过细胞内细胞因子染色进一步验证了这一发现，突出了 3D 单细胞微环境的重要性。这个新的单细胞仿生平台可以很容易地适应其他三维细胞培养支架或其他单细胞检测方法，并且可能成为一种广泛适用的三维单细胞分析系统，用于研究微环境条件对体外细胞功能异质性的影响。
