The biopharmaceutical market has witnessed exponential growth and continuous evolution since the first industrial production of animal cell-based therapeutics in the late 1960s. Nevertheless, the quest for improved biomanufacturing processes faces persistent challenges, particularly in achieving high cell density and productivity. The existing technologies have struggled to meet the ever-increasing global demand for therapeutic agents and traditional bioreactor cultures of animal cells in suspension or as monolayers remain limited. This project originated from a collaborative effort between the academic platform 3D.Fab (ICBMS – Université Lyon 1) and the industrial company Sartorius. Our goal is to leverage the combined scientific expertise of both parties to introduce an innovative biomanufacturing strategy for producing recombinant proteins. We propose the application of bioprinting as a novel biofabrication tool to create complex, controlled, and reproducible 3D adherent cultures of various production cell lines. The objective is to explore the potential of 3D adherent cell cultures in addressing the limitations of current culture-based production. Our work has provided a deeper understanding of the global changes in cellular kinetics within 3D cultures and has described both the advantages and limitations of this culture method for bioproduction. We demonstrated that mAb-producing cell lines cultivated in such 3D bioprinted constructs exhibited higher specific productivity than our suspended bioreactor culture reference, and equivalent specific productivity compared to values reported in the literature for optimized fed-batch suspension cultures. Thus, we have demonstrated the potential of this technology to increase cell-specific productivity and raised questions about its potential to surpass the current maximum cell-specific productivity of 90 pg/cell/day. The 3D bioprinted culture of these production cell lines could be scaled up and cultivated in a custom bioreactor system with full control over key culture parameters. However, the inability of 3D cell culture to reach the high cell densities achieved in standard culture processes results in low volumetric productivity that cannot compete with current industrial bioprocesses. Our work has demonstrated that optimizing bioprinting parameters and hydrogel properties could enhance cell proliferation and increase the maximum cell density achievable. Nonetheless, while it is theoretically possible for cells to grow at very high cell density by colonizing 100% of the construct volume, it remains uncertain whether such a phenomenon is practically achievable. In fact, the greater resemblance of 3D cell culture to the in vivo environment may also trigger in vivo-like responses, such as cell-contact-induced apoptosis. Our research has indicated the presence of successive growth phases interspersed with stationary phases or even a reduction in cell density for some cell lines, which aligns with the growth-apoptosis cycles observed in tissues. Furthermore, existing in situ transfection tools have been studies for progressive insertion of genetic material in cells grown in 3D cultures. Such technology has not yet achieved efficiencies equivalent to those reported in monolayer or suspension cultures. Our work has not been able to overcome this obstacle, but we have proposed suggestions for improving in situ transfection efficiency. The insight provides by this work led to the conclusion that, with current technical capabilities, this technology does not present a compelling alternative to standard large-scale industrial bioproduction processes for recombinant proteins. Nevertheless, its similarity to the in vivo environment, increased specific productivity, and the cell-protective effects of the hydrogel matrix against hydrodynamic stresses present intriguing potential for personalized or niche therapies, such as stem cell-based treatments or extracellular vesicle production.