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

Published: April 30, 2023

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

開示

The authors have nothing to disclose.

Materials

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

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記事を引用
3-Dimensional Culture of Lung Carcinoma Cells: A Method To Study Cell-Matrix Interactions. J. Vis. Exp. (Pending Publication), e20234, doi: (2023).

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