Magnetically Bioprinted 3D Spheroid Co-cultures: A Method for Co-culturing Cancerous Cells and Fibroblasts

Abstract

Source: Noel P. et al. Preparation and Metabolic Assay of 3-dimensional Spheroid Co-cultures of Pancreatic Cancer Cells and Fibroblasts. J. Vis. Exp. (2017)

This video explains the 3D magnetic bioprinting of co-cultured pancreatic cancer cells and fibroblasts. Pancreatic cancer cells and activated pancreatic fibroblast cells are co-cultured and magnetized using a biocompatible nanoparticle assembly, to generate spheroids, under the influence of magnetic forces.

Protocol

1.Culture of Pancreatic Cancer 3D Spheroids using Magnetic Bioprinting Using standard aseptic tissue culture technique, culture cells of interest in a T75 flask to a confluency of 70-80% in appropriate growth media. NOTE: Typically, 5-7 × 106 cells from a 70-80% confluent T75 flask were harvested. Two different cell types were used in this study – Patu8902 (pancreatic tumor cells) and PS1 cells (activated pancreatic stellate cells). Both cell lines were cultured in…

Materials

0.05% Trypsin EDTA	Sigma	25300-054
96 well cell repellant plate	USA Scientific/ Griener Bio-One	655970	spheroid growth plate
Cellometry Cell counting slides	Nexcelom	CHT4-SD100-002
n3D Magnetic Spheroid drive	Nano3D Bioscience	655841	Part of Spheroid Bioprinting Kit
n3D Magnetic Spheroid holding drive	Nano3D Bioscience	655841	Part of Spheroid Bioprinting Kit
n3D NanoShuttle-PL	Nano3D Bioscience	655841	Part of Spheroid Bioprinting Kit: store at 4 °C
Patu8902 cells	Laboratory Stock		pancreatic ductal adenocarcinoma cells
PS1 cells	Laboratory Stock		activated pancreatic stellate cells
RPMI1640	Gibco	11875-093	growth media for cells, spheroids