Bioverse | 1 Cell Type + 405nm

Design of Blueprint

Software used to draw 3D model

Name of software

BioCAD

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

Spacing between lines

Line space = 2mm

Bioinks

Biomaterial used

Company supplying biomaterial

Composition

Formulation of bioink

Any bioink modified with methacryloyl substitution groups, which crosslinks in the presence of the LAP photoinitiator (lithium acylphosphinate salt) and exposure to 405nm blue light. Prior to bioprinting, the photo-crosslinkable bioink can mixed with a high concentration of cells with our CELLMIXER, for a one-step bioprinting process.

Cells

Cell type(s)

Use your desired cell type

Cell concentration

Select a desired/relevant cell concentration for your experiments
Ccell (cells/ml) =

Calculate the amount of bioink needed for your experiments, Vbioink.
For example, the volume of bioink per construct is 100µL. If you need to print 30 constructs, then the bioink volume needed is 3mL.
Vbioink = Vconstruct × Nconstructs

Calculate the amount of cells needed.
Ncells = Ccell (1.1×Vbioink).

Resuspend the cells (Ncells) in 1/10th the volume of the bioink.
Vcell suspension = 0.1×Vbioink
For example, if Vbioink = 3mL, then Vcell suspension = 0.1×3mL = 0.3mL

Mix the cell suspension with the bioink using the CELLMIXER, following the application note, to obtain a homogenous cell distribution. Alternatively, you can use a sterile micro-spatula to mix the cells and bioink and then load the cell-laden bioink into a 3cc cartridge.

Place the cartridge (3cc), loaded with the cell-laden bioink, in the bioprinter (Printhead 1)

Culture media composition

Use the required growth and differentiation media for your cell type

Bioprinter

Model and Manufacturer

INKREDIBLE 3D Bioprinter by CELLINK

Bioprinting Technology & parameters

Pneumatic-driven microextrusion

Pressure

Nozzle type and size for PH1: Conical tip, 250µm ID (25 GA)
Printing pressure for PH1: 25-35 kPa

Printing speed

600 mm/min

Printhead temperature

Room temperature (22°C)

Printbed temperature

Room temperature (22°C)

Crosslinking process of bioink

During the bioprinting process, the 405nm blue light curing system will crosslink each layer, following the same printing path. The default speed is 600 mm/min. If needed, the speed can be decreased by turning the control knob counter-clockwise in order to increase the crosslinking time for each layer.
Once the bioprinting process is completed, transfer the constructs to multiple wellplates (eg. 24 wellplates) prefilled with culture medium (eg. 1mL per well) and incubate in standard culture conditions (37°C, 5% CO2 and 95% relative humidity). Change the media every second day or as often as needed.

Crosslinking method

UV

Bioprinting metrics

Time for bioprinting and crosslinking process per construct

37 seconds per construct

Volume of bioink per construct

0.10ml

Post-bioprinting

Time of 3D culture in vitro

Culture the 3D bioprinted cell-laden constructs for the amount of days that you wish to evaluate. If you’re interested in evaluating cell viability, a 14-day study is recommended. If you want to evaluate the extracellular matrix production or cell differentiation, we recommend at minimum a 28- or 42-day in vitro study.

Stability of constructs in vitro?

During the in vitro 3D culture, you can analyze cell viability, morphology proliferation as well as production of ECM components and gene expression.

1 Cell Type + 405nm

Protocol