Bavli et al. (2021) A highly sensitive analysis platform for the characterization of 3D-cultured single-cell-derived clones. Developmental Cell 56, 1–14. (Link)

Single cells are encapsulated in 3-D Life Hydrogel spheres via an oil-phase ‘‘pinching’’ process in a microfluidic device and subsequently cultured in multiwell plates to generate clones of cells. To culture mouse embryonic stem cells (mESCs) in these spheres the hydrogel components were mixed with several additives including gelatin. mESCs maintained pluripotency over at least 8 days of culture indicating the suitability of this 3D culturing system to promote stemness of ESCs even without pluripotent media. Application: single cell analysis. Cells Used: PC9 (lung adenocarcinoma, human), ZHBTc4 ESCs (mouse embryonic stem cells). Methods used: microsphere production, microsphere cultivation, scRNA-seq, fluorescent labeling of hydrogel via biotin-streptavidin attachment.