3D Co-culture of Lung Cancer Cells with CAFs: An In Vitro Model System to Study Tumor Progression

- In 3D co-cultures, multiple cell types are grown together in matrices like a basement membrane matrix, that mimics the natural extracellular matrix microenvironment, facilitating cell-cell and cell-matrix interactions. To establish an embedded 3D co-culture prepare a uniform suspension of cancer-associated fibroblasts or CAFs and lung cancer cells taken in two is to one ratio, in the basement membrane matrix. Maintain the suspension on ice to prevent matrix solidification at ambient temperatures.

Transfer an appropriate volume of suspension into a cell culture plate. Incubate at 37 degrees Celsius for the required duration to co-embed the cells in the matrix. To establish an overlaid 3D co-culture, prepare a suspension of lung cancer cells in basement membrane matrix, in the desired cell density. Maintain the suspension on ice. Pipette an appropriate volume of this cell-matrix suspension into a cell culture plate. Incubate at 37 degrees Celsius to set up the embedded cancer cell monoculture.

Subsequently transfer the desired volume of CAFs suspended in preferred culture media, over the embedded cancer cells. In 3D co-cultures, CAFs enhance cancer cells spheroid formation and attr