Macrophage secretion heterogeneity in engineered microenvironments revealed using a microwell platform

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 tec...

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Bibliographic Details
Published in:Integrative biology (Cambridge) 2016-07, Vol.8 (7), p.751-76
Main Authors: McWhorter, Frances Y, Smith, Tim D, Luu, Thuy U, Rahim, Maha K, Haun, Jered B, Liu, Wendy F
Format: Article
Language:English
Subjects:
Animals
Cell Adhesion - physiology
Cell Separation - instrumentation
Cell Size
Cells, Cultured
Cellular Microenvironment - physiology
Cytokines - secretion
Equipment Design
Female
Immunoassay - instrumentation
Lab-On-A-Chip Devices
Macrophages - cytology
Macrophages - secretion
Mice
Mice, Inbred C57BL
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Summary: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. A microwell system for detection of secreted products from adherent cells is used to demonstrate that macrophage adhesive context and cell shape regulate cytokine secretion and population heterogeneity.
ISSN:1757-9694
1757-9708
DOI:10.1039/c6ib00053c