3D Co-culture of Lung Cancer Cells with CAFs: An In Vitro Model System to Study Tumor Progression
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 two invitro model systems to capture the tissue architectural changes during lung squamous carcinoma progression in a 3-dimensional (3D) co-culture with cancer-associated fibroblasts. This system provides a unique platform to investigate the role of intrinsic and extrinsic changes that affect the tumor phenotype.
Protocol
1.Passaging and Culturing TUM622 Cells and CAFs in 2D Cultures Passaging and culturing TUM622 cells 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 b…
Açıklamalar
The authors have nothing to disclose.
Materials
| CoolRack CFT30 | Biocision | BCS-138 | For 3D culture |
| CoolSink XT96F | Biocision | BCS-536 | For 3D culture |
| Cultrex (preferred for co-culture) | Bio-Techne | 3443-005-01 | For 3D culture |
| Lab-Tec II chambered #1.5 German Coverglass System | Nalge Nunc International | 155379 (2) | For 3D culture |
| Lab-Tec II chambered #1.5 German Coverglass System | Nalge Nunc International | 155409 (8) | For 3D culture |
| L-Glutamine | Gibco | 25030-081 | For CAFs |
| Matrigel (preferred for monoculture) | Corning | 356231 | For 3D culture |
| ReagentPack Subculture Reagents | Lonza | CC-5034 | For TUM622 cell dissociation |
| Pen/Strep | Gibco | 15140-122 | For CAFs |
| Heat-inactivated FBS | Gibco | 10082-147 | For CAFs |
| TrypLE Express | Gibco | 12604-021 | For CAF dissociation |
| Cell Strainer 40um | ThermoFisher | 352340 | For passing TUM622 cells |
| RPMI | ThermoFisher | 11875-093 | For CAFs |
Bu Makaleden Alıntı Yapın
3D Co-culture of Lung Cancer Cells with CAFs: An In Vitro Model System to Study Tumor Progression. J. Vis. Exp. (Pending Publication), e20235, doi: (2023).