A Method for 3D Bioprinting Murine Cortical Astrocytes

Take an astrocyte suspension.

Add a bioink solution containing gelatin, photocrosslinkable gelatin derivatives, photoinitiators, fibrinogen, and laminin.

Transfer the suspension to a syringe with a blunt needle.

Chill the syringe for bioink gelling. Connect the syringe to the bioprinter's printhead.

Position the needle appropriately to the culture plate. Initiate bioprinting with set parameters and the design.

Astrocyte-containing bioink is pushed out of the needle, and is deposited in layers onto the plate, forming a three-dimensional structure with trapped astrocytes.

Expose the bioprinted construct to ultraviolet light. The photoinitiator aids in gelatin derivative crosslinking, enhancing bioconstruct stability.

Treat with thrombin and calcium ions.

Thrombin, with calcium ions, converts fibrinogen into fibrin, assembling into a stable 3D biopolymer network.

Discard the solution and wash with buffer to remove the crosslinking agents.

Add astrocyte media and incubate. Laminin aids astrocyte adhesion and spreading, forming a 3D neural-like tissue rich in astrocytes.