Macrophage secretion heterogeneity in engineered microenvironments revealed using a microwell platform.
Macrophage secretion heterogeneity in engineered microenvironments revealed using a microwell platform.
复制标题
使用微孔平台揭示工程微环境中巨噬细胞分泌的异质性。
DOI：
10.1039/c6ib00053c
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Liu,WendyF
中科院分区：
文献类型：
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作者：
McWhorter,FrancesY;Smith,TimD;Luu,ThuyU;Rahim,MahaK;Haun,JeredB;Liu,WendyF
分泌蛋白在协调细胞群的反应中发挥着重要作用。然而，在单细胞水平上对蛋白质分泌响应微环境线索的动态变化的定量理解仍然难以捉摸。使用传统分子技术进行的测量通常需要大量培养，因此无法捕获细胞群内的多样性。基于芯片的技术的最新进展表明，单细胞测量可以为细胞激活和功能的时间动态提供重要的见解，但这些工具对粘附细胞微环境的控制有限。在这里，我们创建了一个单细胞细胞因子检测平台，可以控制物理和粘附微环境。我们通过检查暴露于不同剂量的可溶性刺激和不同粘附基质上的巨噬细胞的细胞因子分泌来验证该平台。我们还使用该平台证明细胞形状影响单个巨噬细胞细胞因子的分泌。总之，这些结果表明微孔系统能够在受控粘附环境中检测单个巨噬细胞分泌的细胞因子。该技术可广泛应用于检测任何贴壁细胞类型的分泌产物。
Secreted proteins play a major role in orchestrating the response of cell populations. However, a quantitative understanding of the dynamic changes in protein secretion in response to microenvironmental cues at the single cell level remains elusive. Measurements taken using traditional molecular techniques typically require bulk cultures, and therefore cannot capture the diversity within cell populations. Recent advances in chip-based technologies have shown that single cell measurements can provide important insights into the temporal dynamics of cellular activation and function, but these tools have had limited control of the adhesive cellular microenvironment. Here, we created a single cell cytokine detection platform that allows for controlled physical and adhesive microenvironment. We validated the platform by examining cytokine secretion of macrophages exposed to varying dosages of soluble stimulation and on different adhesive substrates. We also used the platform to demonstrate that cell shape affects single macrophage cytokine secretion. Together, these results show the ability of the microwell system to detect secreted cytokines from individual macrophages in controlled adhesive environments. This technique may be broadly applied to detect secreted products from any adherent cell type.