3-Dimensional Culture of Lung Carcinoma Cells: A Method To Study Cell-Matrix Interactions

- First, thaw vials of basement membrane matrix at 4 degrees Celsius overnight. The basement membrane matrix consists of proteins present in the tumor microenvironment that help maintain cancer cells' characteristics in vitro. The next day, place the basement membrane matrix vial on ice to retain the matrix in liquid state. Take lung cancer cells into a pre-cooled centrifuge tube and centrifuge at 4 degrees Celsius. Aspirate the supernatant and gently tap to dislodge the pellet and place it on ice. Next, add an appropriate volume of basement membrane mix into this tube and mix to make a uniform cell suspension. Perform all steps at low temperature to prevent the basement membrane from gelling.

Transfer an appropriate volume of cell matrix suspension to the center of each well in a precooled well plate, covering the entire well. Incubate the well plate for the matrix to solidify. Examine under a microscope to ensure even distribution of cells within the matrix. Add 3D culture complete medium into each well and incubate at 37 degrees Celsius. This medium facilitates cell matrix interactions. Thus, the cells form organoids with acinar-like structures. In the following protocol, we wil