Astrocytes, the most abundant glial cells in the central nervous system, play a critical role in supporting the normal physiological function of neurons. Recent studies have revealed that astrocyte transplantation can promote axonal regeneration and functional recovery after spinal cord injury. Biomaterial can be designed as a growth-permissive substrate and serve as a carrier for astrocyte transplantation into injured spinal cord. In this study, we developed a method to generate collagen microspheres encapsulating astrocytes by injecting the mixture of collagen and astrocytes into the cell culture medium with a syringe controlled by a syringe pump.