Extrusion bioprinting is a promising tool for autonomous medical care and biomanufacturing during long-duration crewed space missions. However, the implementation of this technology depends on how cell-laden bioinks can be brought to the space platform without compromising their biological functionality. In this study, a supply chain solution is introduced in which cells and acellular biomaterial ink are transported separately (cells as cryopreserved vials at −80°C and the ink at 4°C) and are combined into the final bioink minutes before printing. This cryo-based bioink approach is validated against a freshly prepared reference bioink and a premixed, cold-stored bioink using a parabolic flight campaign, including the logistic chain of road transport to the flight site, crew-level preparation in Bordeaux, in-flight printing during microgravity, and post-flight sample return to Dresden for cultivation. Human dermal fibroblasts printed with the cryo-based bioink reach viabilities above 90%, comparable to the reference group. In contrast, the viability of the cells printed from cold-stored bioink declined and with no detectable cells at the final cultivation time point. Our results show that decoupling cells and biomaterial ink through cryopreservation preserves biological functionality and enables a scalable bioink supply chain using infrastructure available on crewed space platforms.