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
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).