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…
Disclosures
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