Cryobioprinted human tumor models with shelf-stable programmability.

Area of Science:

• Bioprinting and Tissue Engineering
• Cryobiology
• Cancer Modeling

Background:

• Cryobioprinting offers simultaneous fabrication and cryopreservation of tissue analogs.
• Effective cryopreservation of living constructs is limited by the need for cell-type and bioink-specific conditions.
• Developing shelf-ready tissue models is crucial for advancing drug screening platforms.

Purpose of the Study:

• To formulate a decellularized extracellular matrix (dECM)-based hydrogel bioink with cryoprotective agents (CPAs) for creating shelf-ready pancreatic cancer models.
• To discover an optimal CPA combination for cryobioprinting tumor-stroma constructs.
• To evaluate the viability and metabolic activity of cryopreserved constructs for in vitro applications.

Main Methods:

• Combinatorial screening of CPAs within a dECM hydrogel.
• Development of a novel melezitose-glycerol-dECM formulation.
• Exometabolomics analysis to assess metabolic activity post-thawing.
• Assessment of cell viability and suitability for in vitro drug screening.

Main Results:

• A novel melezitose-glycerol-dECM formulation demonstrated superior cryoprotective properties for both tumor and stroma compartments.
• Cryopreserved constructs maintained metabolic activity comparable to non-frozen controls 14 days post-thawing.
• Cryobioprinted tumor-stroma models exhibited increased cell viability and were suitable for in vitro drug screening.

Conclusions:

• The optimized cryoprotective strategy using dECM-CPA bioinks enables the creation of shelf-ready, living tissue constructs.
• This approach enhances cell viability and metabolic function after cryopreservation.
• The developed method opens avenues for cryobioprinting diverse tissue types for widespread drug screening and regenerative medicine.