Bioprinting Spheroids/Organoids Using Custom G-code

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Bioprinting Spheroids/Organoids Using Custom G-code

Overview

Spheroids and organoids are 3D culture models that have served as simple, yet physiologically relevant organ samples when compared to 2D cell cultures. Researchers have used these models as a platform to run screening experiments, measuring different markers relevant to the tissue type they are testing. This protocol runs you through the process of bioprinting spheroids/organoids using custom g-code on the Allevi system – a practical way to deposit cell-laden hydrogels and media for spheroid or organoid creation.

Materials

Methods: Bioprinting Spheroids/Organoids Using Custom G-code

  1. Create your G-code file by using the following template:
1  G92 E0 
2
3  ;dot 14  T_; set printing extruder (T0 = extruder 1, T1 = extruder 2, T2 = extruder 3)
5  G1 X__ Y__ Z__ F600; move to position (__, __, __) at speed of 600 mm/min without 6  extruding material
7  G1 X__ Y__ E0.1; begin extruding material (repeat same coordinates as above)
8  M400
9  G4 S_; set extrusion time
10 G92 E0; stop extrusion
11 G1 X__ Y__ Z__; move extruder to the next desired position

Note: Allevi 2 users may need to enter M106 T_ as well to turn the extruder on. Similarly, they may need to enter M107 T_ to turn the extruder off.

  1. Copy and paste lines 3-10 to make more dots. Just ensure to change each coordinate, extruder and extrusion time appropriately;
  2. Save your file as a .gcode file;
  3. Add this G-code to your dashboard and print!

We hope you found this protocol helpful for bioprinting spheroids/organoids using custom g-code! You’re now ready to image and analyze your 3D bioprinted structures. Click here for a full list of analysis protocols.

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