Rapid Production of Cell-Laden Microspheres Using a Flexible Microfluidic Encapsulation Platform.

Abstract

This study establishes a novel microfluidic platform for rapid encapsulation of cells at high densities in photocrosslinkable microspherical hydrogels including poly(ethylene glycol)-diacrylate, poly(ethylene glycol)-fibrinogen, and gelatin methacrylate. Cell-laden hydrogel microspheres are advantageous for many applications from drug screening to regenerative medicine. Employing microfluidic systems is considered the most efficient method for scale-up production of uniform microspheres. However, existing platforms have been constrained by traditional microfabrication techniques for device fabrication, restricting microsphere diameter to below 200 µm and making iterative design changes time-consuming and costly. Using a new molding technique, the microfluidic device employs a modified T-junction design with readily adjustable channel sizes, enabling production of highly uniform microspheres with cell densities (10-60 million cells mL-1 ) and a wide range of diameters (300-1100 µm), which are critical for realizing downstream applications, through rapid photocrosslinking (≈1 s per microsphere). Multiple cell types are encapsulated at rates of up to 1 million cells per min, are evenly distributed throughout the microspheres, and maintain high viability and appropriate cellular activities in long-term culture. This microfluidic encapsulation platform is a valuable and readily adoptable tool for numerous applications, including supporting injectable cell therapy, bioreactor-based cell expansion and differentiation, and high throughput tissue sphere-based drug testing assays.