High‐Throughput Measurements of Intra‐Cellular and Secreted Cytokine from Single Spheroids Using Anchored Microfluidic Droplets

Abstract

communication and help regulate the function on the scale of whole organs and organism. The different types of molecules are present at different concentrations in vivo and act over a range of time scales, from seconds to days. Indeed, variations in the rate of secretion of different molecules indicate the state of the cells, for example, as a response to changes in their environment. Standard methods for measuring secreted molecules in typical cell culture experiments, however, require the presence of a large number of molecules, which translates into volumes of tens to hundreds of µL and give information at the level of large populations of cells. As a result, these techniques hide the heterogeneities that may exist on the scale of individual cells. It is becoming increasingly clear, however, that these heterogeneities are present and play a determining role in many biological processes, such as the immune response to an unknown pathogen, [1] or as a prognostic marker for some cancer types. [2] 2002303 (2 of 11)

Cell3: A new vision for study of the endomembrane system in mammalian cells

“…The first population contained the cells to form the spheroids and the second population of droplets was used to manipulate the spheroids, for example allowing establishment of co-cultures or treatment with drug libraries [48]. This device has also been used to study the formation of organoids by fluorescence imaging [49], and to study the secretion of vascular endothelial growth factor by mesenchymal stem cell spheroids [50]. These studies clearly show the potential of this method to grow and manipulate spheroids, and applications for study of the endomembrane system in spheroids grown in such a device should be extensive.…”

Cell3: a new vision for study of the endomembrane system in mammalian cells

“…The first population contained the cells to form the spheroids and the second population of droplets was used to manipulate the spheroids, for example allowing establishment of co-cultures or treatment with drug libraries [ 48 ]. This device has also been used to study the formation of organoids by fluorescence imaging [ 49 ], and to study the secretion of vascular endothelial growth factor by mesenchymal stem cell spheroids [ 50 ]. These studies clearly show the potential of this method to grow and manipulate spheroids, and applications for study of the endomembrane system in spheroids grown in such a device should be extensive.…”

Technological Advances in Tumor-On-Chip Technology: From Bench to Bedside

“…Droplet microfluidic platforms have been developed to facilitate the generation of multicellular tumor spheroids by mixing and encapsulating cells in a single droplet, which is protected by an immiscible liquid phase [42][43][44]. Taking advantage of a high surface-area-to-volume ratio, these platforms offer the possibility of high-throughput and homogeneous spheroid formation [45].…”

“…Nonetheless, the capabilities of microfluidic technologies can also be coupled with imaging tools to unravel the molecular behavior of spheroids. As an example, Saint-Sardos and colleagues recently developed their droplet microfluidic platform to enable the measurement of both intracellular and secreted cytokines from spheroids [45]. A secondary droplet, containing anti-VEGF-A beads and anti-VEGF-A secondary fluorescent antibody, was delivered and anchored next to the spheroid droplets.…”