A High-Throughput Automated Microfluidic Platform for Calcium Imaging of Taste Sensing.

摘要: The human enteroendocrine L cell line NCI-H716, expressing taste receptors and signaling elements, constitutes a unique model for the studies of cellular responses to glucose, appetite regulation, gastrointestinal motility, insulin secretion. Targeting these gut may provide novel treatments diabetes obesity. However, NCI-H716 cells are cultured in suspension tend form multicellular aggregates, preventing high-throughput calcium imaging due interferences caused by laborious immobilization stimulus delivery procedures. Here, we have developed an automated microfluidic platform that is capable trapping more than 500 single into microwells with loading efficiency 77% within two minutes, delivering multiple chemical stimuli performing enhanced spatial temporal resolutions when compared bath perfusion systems. Results revealed presence heterogeneity type, concentration, order applied sweet bitter stimuli. Sucralose denatonium benzoate elicited robust increases intracellular Ca(2+) concentration. glucose evoked rapid elevation followed reduced subsequent stimulation. Using Gymnema sylvestre as blocking agent receptor confirmed different were utilized tastes. This cost-effective, easy fabricate operate, be generally applicable high-content single-cell analysis drug screening.