3-Dimensional Culture of Lung Carcinoma Cells: A Method To Study Cell-Matrix Interactions

Abstract

Source: Chen, S. et. al. Multidimensional Coculture System to Model Lung Squamous Carcinoma Progression. J. Vis. Exp. (2020).

This video describes the technique of 3-D culture of lung carcinoma cells to study the cell-matrix interactions. In the protocol, we will perform the 3D culture of TUM622 lung carcinoma cells to explore its potential of forming organoids in vitro.

Protocol

1. Passaging and Culturing TUM622 Cells in 2D Cultures

1. Warm 3D culture medium and cell dissociation reagents (see Table of Materials) for TUM622 cells at 37 °C.
2. Passage TUM622 cells at 80% confluency in 2D flasks. Usually, this occurs 1 week after passaging.
3. Discard old medium from a T75 flask and wash once with 6 mL of HEPES buffer. Avoid pipetting directly onto the cells.
4. Aspirate the HEPES buffer. Add 4 mL of trypsin/EDTA (0.25 mg/mL, see Table of Materials) for a quick rinse and discard the trypsin/EDTA.
5. Add 2 mL of trypsin/EDTA and incubate at 37 °C for 5 min....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Bronchial Epithelial Growth Medium	Lonza	CC-3170 BEGM
Cell Strainer 40um	ThermoFisher	352340	For passing TUM622 cells
CoolRack CFT30	Biocision	BCS-138	For 3D culture
CoolSink XT96F	Biocision	BCS-536	For 3D culture
Cultrex 3D Cell Harvesting Kit	Bio-Techne	3448-020-K
Matrigel (preferred for monoculture)	Corning	356231	For 3D culture
Lab-Tec II chambered #1.5 German Coverglass System	Nalge Nunc International	155409 (8)	For 3D culture
Lab-Tec II chambered #1.5 German Coverglass System	Nalge Nunc International	155379 (2)	For 3D culture
ReagentPack Subculture Reagents	Lonza	CC-5034	For TUM622 cell dissociation