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Updated: Sep 15, 2026

Preparation of Cell Extracts by Cryogrinding in an Automated Freezer Mill
Published on: January 29, 2021
Temperature Controlled Cryoprinting
Leo Lou1,2, Linnea Warburton1,3, Boris Rubinsky4,5
1Department of Mechanical Engineering and Department of Bioengineering, University of California Berkeley, Berkeley, CA 94720, USA.
Abstract:
This chapter reviews the development of temperature-controlled cryoprinting (TCC), a bioprinting approach that integrates extrusion with controlled freezing to stabilize constructs during fabrication. Conventional extrusion printing struggles to balance ink viscosity, mechanical fidelity, and biological compatibility, often necessitating secondary stabilization steps that compromise structural integrity or viability. TCC overcomes these limitations by freezing each voxel under regulated thermal conditions, maintaining structural stability throughout the printing process and eliminating the need for post-print freezing. The review traces the conceptual origins of TCC from early cryobiology research on ice physics and cell survival, through proof-of-concept demonstrations of cryogenic 3D printing, to the progressive refinement of methods including multi-layer cryolithography, freezing-modulated crosslinking, and modular cryoprinting systems. It summarizes material innovations such as alginate-agar inks and explores how freezing history influences diffusion and microstructural anisotropy. Applications are presented in tissue engineering, where TCC supports cryopreservation of cell-laden scaffolds and vascularizable tissues, and in food engineering, where it enables production of customized frozen foods with reproducible textures. The intellectual property framework that has developed alongside the research is also reviewed. Together, these studies establish TCC as an approach that unites fabrication and preservation, with implications for both biomedical and food applications.

