Ref No: PPR-TP60505
Approval date: 02-JAN-2023
Author: PT, AB, VK. Version: 2

PRINTING PROTOCOLPCLThis is a suggested procedure, please adjust according to your experimental needs.
To maintain the sterility of the product, work under sterile conditions.

Protocol aim

Materials needed

The aim of this protocol is to provide instructions forprinting biocompatible and biodegradable 3D
structures with CELLINK’s polycaprolactone (PCL).
PCL can be extruded through both heating andmixing with a solvent. This protocol has been
optimized for heat extrusion at 180°C using theThermoplastic printhead and the BIO X but can also
be extruded using the aluminum cartridge and tips
with the INKREDIBLE+.

••••

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PCL*
Thermoplastic printhead*
BIO X*
Petri dish**The product can be purchased in the CELLINKshop at www.cellink.com/shop.

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Protocol

1.

Filling cartridge with PCL

MATERIAL
PCL
Thermoplastic printhead

DESCRIPTION•

Load the cartridge as recommended in the Thermoplastic Printhead User Manual.

Note: To ensure the most efficient heating of PCL, fill maximum half of the cartridge with PCL.
Note: If mixing with a solvent, ensure that the solvent is compatible with the Thermoplastic printhead (BIO X)
or the stainless-steel cartridge, plunger, and the tips (INKREDIBLE+).

2.

Pre-heatingMATERIALBIO X

DESCRIPTION•

Attach the Thermoplastic printhead to the BIO X and start pre-heating at 180°C. Heat for 15 min until thePCL is melted.

3.

Printing

MATERIALBIO XPetri dish

DESCRIPTION•

Set layer height at 80% of the nozzle diameter. Start with a printing temperature of 180°C and thepressures showed in Figure 1.

Note: Due to the need to fuse filaments of successive layers together, it is recommended to use a layerheight that is smaller than the nozzle diameter. This is to allow the filaments to fuse and to account for
shrinkage of the layers during the cooling process.
Note: If PCL is extruding inconsistently, the tip may be dirty. Wipe the tip with steel wool to remove theexcess polymer. If the PCL is turning brown, the temperature is too high. Lower the temperature or replace
the PCL in the cartridge. Re-start from step 1, then flush the nozzle with new PCL from the printer overview
page or the utilities menu. If necessary, run a calibration script to reoptimize the printing characteristics.

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Figure 1. PCL filament thickness dependency on printing speed and pressure when extruded at 180°C.

4.

Solidification

DESCRIPTION•Let PCL cool down and solidify after printing. Do not touch the PCL construct until it is completely cooled
as this can deform the structure.

Note: The temperature-controlled print bed on the BIO X can be used to accelerate or slow down this coolingprocess. Sintering of PCL filaments is possible. If a PCL/solvent mixture is printed, make sure that theresulting construct is placed in a well-ventilated area for drying to ensure the solvent is removed prior to cellseeding.
Note: If the structure contracts inwards during the printing process, there may be a need for additionalcooling between successive layers. Modify the G-code to pause the print for 1 min between layers to allow
solidification of already deposited material.

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