High-throughput measurements of intra-cellular and secreted cytokine from single spheroids using anchored microfluidic droplets

摘要: Abstract While many single-cell approaches have been developed to measure secretion from anchorage-independent cells, these protocols cannot be applied adherent especially when cells requires cultured in 3D formats. Here we demonstrate a platform the secretions individual spheroids of human mesenchymal stem within microfluidic droplets. The allows us quantify hundreds each device, by using secondary droplet bring functionalized micro-beads into proximity with spheroid. We focus on vascular endothelial growth factor (VEGF) and distribution levels that presents broad heterogeneity population spheroids. Moreover, intra-cellular level VEGF-A spheroid, measured through immuno-staining, correlates well extra-cellular measurement, indicating heterogeneities observed at result variations scale cells. Finally, model molecular accumulation droplets find physical confinement is crucial for measurements protein secretions. predicts time achieve which scales volume. Therefore first provide several new biological insights.