Protocols

Blood Vessel

Protocol

Design of Blueprint

Software used to draw 3D model

Name of software

Autodesk® 123D Design

Dimensions of 3D model

Tube geometry: 6mm x 10mm (ODxH)

Software used to convert 3D model to bioprinting protocol and toolpath

Name of software

b.i. Slic3r (v1.2.9)

Spacing between lines

Layer height = 0.30mm

Porosity of model

External perimeters extrusion width = 0.30mm

Layer thickness

Printing speed, F = 600mm/min

Bioinks

Biomaterial used

CELLINK Bioink

Company supplying biomaterial

CELLINK

Composition

Formulation of bioink

Nanocellulose and sodium alginate-based bioink

References

b.i. 3D Bioprinting Human Chondrocytes with Nanocellulose–Alginate Bioink for Cartilage Tissue Engineering Applications. Kajsa Markstedt, Athanasios Mantas, Ivan Tournier, Héctor Martínez Ávila, Daniel Hägg, and Paul Gatenholm. Biomacromolecules 2015 16 (5), 1489-1496.
dx.doi.org/10.1021/acs.biomac.5b00188

3D bioprinting of human chondrocyte-laden nanocellulose hydrogels for patient-specific auricular cartilage regeneration. Héctor Martínez Ávila, Silke Schwarz, Nicole Rotter, Paul Gatenholm. Bioprinting 2016, Volumes 1–2, 22-35.
dx.doi.org/10.1016/j.bprint.2016.08.003

Cells

Cell type(s)

Human smooth muscle cells (SMC)

Cell concentration

10x106 cells/ml
A suspension of 33x106 SMCs in a volume of 300µL were mixed with 3mL of CELLINK bioink using the CELLMIXER at a ratio of 10:1 (Bioink to cell supension) to obtain a final concentration of 10x106 cells/ml.

Culture media composition

See reference http://dx.doi.org/10.1016/j.biomaterials.2005.10.026

Bioprinter

Model and Manufacturer

INKRIBLE 3D Bioprinter by CELLINK

Bioprinting Technology & parameters

Pneumatic-driven microextrusion

Pressure

15-17 kPa

Printing speed

600 mm/min

Printhead temperature

Room temperature (22°C)

Printbed temperature

Room temperature (22°C)

Crosslinking process of bioink

After the bioprinting process, the cell-laden constructs were crosslinked by submerging in an ionic solution of 100mM CaCl2 for 6 minutes. The constructs were then rinsed and incubated in culture medium in standard culture conditions (37°C, 5% CO2 and 95% relative humidity).

Crosslinking method

Ionic

Concentration

100mM CaCl2

Crosslinking time

6 minutes

Bioprinting metrics

Time for bioprinting and crosslinking process per construct

2 minutes per construct

Volume of bioink per construct

mL

Dimensions of bioprinted construct

OD: 6.1mm, H: 10mm

Post-bioprinting

Time of 3D culture in vitro

7 days

Stability of constructs in vitro?

Macroscopic evaluation of constructs showed no change in shape after 7 days of in vitro culture.

Results

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