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 |
