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  • 3D Printed Hydrogel Multiassay Platforms for Robust Generation of Engineered Contractile Tissues.

3D Printed Hydrogel Multiassay Platforms for Robust Generation of Engineered Contractile Tissues.

Biomacromolecules (2019-12-21)
Rie Kjær Christensen, Christoffer von Halling Laier, Aysel Kiziltay, Sandra Wilson, Niels Bent Larsen
ABSTRACT

We present a method for reproducible manufacture of multiassay platforms with tunable mechanical properties for muscle tissue strip analysis. The platforms result from stereolithographic 3D printing of low protein-binding poly(ethylene glycol) diacrylate (PEGDA) hydrogels. Contractile microtissues have previously been engineered by immobilizing suspended cells in a confined hydrogel matrix with embedded anchoring cantilevers to facilitate muscle tissue strip formation. The 3D shape and mechanical properties of the confinement and the embedded cantilevers are critical for the tissue robustness. High-resolution 3D printing of PEGDA hydrogels offers full design freedom to engineer cantilever stiffness, while minimizing unwanted cell attachment. We demonstrate the applicability by generating suspended muscle tissue strips from C2C12 mouse myoblasts in a compliant fibrin-based hydrogel matrix. The full design freedom allows for new platform geometries that reduce local stress in the matrix and tissue, thus, reducing the risk of tissue fracture.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-α-Actinin (Sarcomeric) antibody produced in mouse, clone EA-53, ascites fluid
Sigma-Aldrich
Quinoline Yellow, Mixture of the mono- and disulfonic acids of Quinoline Yellow
Sigma-Aldrich
Poly(ethylene glycol) diacrylate, average Mn 700
Sigma-Aldrich
Aprotinin from bovine lung, lyophilized powder, 3-8 TIU/mg solid
Sigma-Aldrich
Thrombin from bovine plasma, lyophilized powder, ≥2,000 NIH units/mg protein (E1%/280 = 19.5)
Sigma-Aldrich
Fibrinogen from bovine plasma, Type I-S, 65-85% protein (≥75% of protein is clottable)
Sigma-Aldrich
Propidium iodide, ≥94% (HPLC)
Sigma-Aldrich
Anti-Mouse IgG (H+L), F(ab′)2 fragment, CF488A antibody produced in goat, ~2 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Lithium phenyl-2,4,6-trimethylbenzoylphosphinate, ≥95%
Sigma-Aldrich
Poly(ethylene glycol), BioUltra, 10,000