Acoustic holographic assembly of cell-dense tissue constructs

Area of Science:

• Biotechnology
• Regenerative Medicine
• Tissue Engineering

Background:

• Current biofabrication methods struggle to create large, cell-dense tissue constructs with controlled geometry.
• Physiologically relevant cell densities and centimeter-scale dimensions are crucial for organ-level studies and regenerative medicine.

Purpose of the Study:

• To present an acoustic holographic assembly (AHA) biofabrication scheme for creating cell-dense, centimeter-scale, arbitrarily-shaped tissue constructs.
• To demonstrate the capability of AHA in assembling cells within standard cell culture labware for tissue engineering.

Main Methods:

• Utilized a compact, benchtop acoustic holographic assembly instrument compatible with biolab environments.
• Assembled C2C12 myoblasts in gelatin methacryloyl (GelMA) hydrogel into predefined geometries.
• Cultured assembled constructs under perfusion for five days.

Main Results:

• Rapidly formed large, asymmetric branch-shaped constructs with an average cell density of 40 million cells·ml⁻¹ (local density up to 260 million cells·ml⁻¹).
• Achieved high cell viability of 90.5 ± 4.3% in the assembled constructs.
• Demonstrated single-step assembly of cells into 3D geometries within standard cell culture labware.

Conclusions:

• Acoustic holographic assembly is a promising method for fabricating cell-dense, centimeter-scale tissue constructs with controllable geometry.
• AHA enables the rapid, contactless, label-free assembly of biological cells, overcoming limitations of current biofabrication techniques.
• This technology can facilitate the development of engineered tissues with structural and functional characteristics mirroring native tissues.