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 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.
Department of Biomedical Engineering, University of California Irvine, 2412 Engineering Hall, Irvine, CA 92697, USA. wendy.liu@uci.edu.
Frances Y
FY
McWhorter
Tim D
TD
Smith
Thuy U
TU
Luu
Maha K
MK
Rahim
Jered B
JB
Haun
Wendy F
WF
Liu
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002469:Cell Separation / Q000295:instrumentation*; D048429:Cell Size; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000502:physiology*; D016207:Cytokines / Q000378:metabolism*; D004867:Equipment Design; D005260:Female; D007118:Immunoassay / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL
D016207:Cytokines