Microfluidic switching system for analyzing chemotaxis responses of wortmannin-inhibited HL-60 cells.
The chemotaxis of phosphoinositide kinase-3 (PI3K)-inhibited differentiated HL-60 cells stably expressing CXCR2 was studied in a microfluidic switching gradient device that can generate stable and well-defined forward and reverse gradients. Wortmannin, a widely used PI3K inhibitor, was added during cell preparation and the experiment process. The studies quantify the chemotaxis gradient and the effects of a change in the direction of a CXCL-8 gradient on cell migration. PI3K-inhibited HL-60 cells migrated more efficiently toward the gradient before gradient switching than after, as measured by the effective chemotactic index. The inhibited HL-60 cells also showed that inadequate polarization, slower response time, and reduced cell populations can follow the gradient change. We observed that the role of PI3K in directing cellular response to gradient reversal was important in cell polarization and directional sensing associated with gradient switching.
Liu
Yuxin
Y
Vanderbilt Institute for Integrative Biosystems Research and Education (VIIBRE), Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 37235, USA.
Sai
Jiqing
J
Richmond
Ann
A
Wikswo
John P
JP
D000730:Androstadienes / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects* / Q000502:physiology; D048430:Cell Shape / Q000187:drug effects / Q000502:physiology; D002633:Chemotaxis / Q000187:drug effects*; D016895:Culture Media, Serum-Free; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004867:Equipment Design / Q000295:instrumentation; D018922:HL-60 Cells; D006801:Humans; D044085:Microfluidics / Q000295:instrumentation*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D023063:Receptors, Interleukin-8B / Q000378:metabolism; D012996:Solutions / Q000737:chemistry; D012997:Solvents / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors; D014162:Transfection; D000077191:Wortmannin
D000730:Androstadienes; D016895:Culture Media, Serum-Free; D004791:Enzyme Inhibitors; D000081082:Phosphoinositide-3 Kinase Inhibitors; D023063:Receptors, Interleukin-8B; D012996:Solutions; D012997:Solvents; D000077191:Wortmannin; D004121:Dimethyl Sulfoxide