Cell Microenvironment pH Sensing in 3D Microgels Using Fluorescent Carbon Dots

We report here a 3D cell culture microgel-based system containing carbon dots capable of sensing the pH changes in the cellular microenvironment. We have utilized a simple droplet-based microfluidics methodology for encapsulating cells and fluorescent pH sensitive carbon dots in polyethylene glycol microgels. Since the microfluidics assembly is developed from simple components that can be modified easily to yield microgels of different size, composition, and architecture, it can be utilized to develop complex 3D cell culture scaffolds of desired composition along with spatial control on the polymer composition. The synthesized pH sensitive carbon dots possess green fluorescence emission, which increases as the pH is lowered from neutral to acidic. Since the probe sensitivity to pH change is well within the physiologically relevant range (pH 5.8-7.7), the probe can be used for detecting a lowering of pH as the cells proliferate or undergo various biological processes. We demonstrate that the nanoprobes as well as the process of forming the microgel beads with nanoprobes and mammalian cells is biocompatible, and the cells easily proliferate inside the microgels. The changes in pH as the mammalian cells grow in the microgels is easily monitored via fluorescence microscopy, suggesting that the platform can be used to study time dependent changes in cellular microenvironment pH and can be easily utilized to monitor cellular growth, disease progression, etc.