Cell Culture and Coculture for Oncological Research in Appropriate Microenvironments

Abstract

With the increase in knowledge on the importance of the tumor microenvironment, cell culture models of cancers can be adapted to better recapitulate physiologically relevant situations. Three main microenvironmental factors influence tumor phenotype: the biochemical components that stimulate cells, the fibrous molecules that influence the stiffness of the extracellular matrix, and noncancerous cells like epithelial cells, fibroblasts, endothelial cells, and immune cells. Here we present methods for the culture of carcinomas in the presence of a matrix of specific stiffness, and for the coculture of tumors and fibroblasts as well as epithelial cells in the presence of matrix. Information is provided to help with choice and assessment of the matrix support and in working with serum‐free medium. Using the example of a tissue chip recapitulating the environmental geometry of carcinomas, we also highlight the development of engineered platforms that provide exquisite control of cell culture parameters necessary in research and development. © 2019 by John Wiley & Sons, Inc.

Technological Advances in Tumor-On-Chip Technology: From Bench to Bedside

“…To reconstitute the ECM on-chip, synthetic scaffold-based culturing methods are frequently chosen, because they focus on culturing cell types in a 3D environment that is able to closely mimic natural ECM. Scaffolds can be synthetic, such as poly(ethylene glycol) [55], or naturally derived hydrogel matrices, including Matrigel or collagen I and IV [8,56]. Although these polymers are similar to the ECM components found in solid tumors, their characteristics vary in terms of composition, structure and stiffness, depending on their method of production [6].…”

“…For example, a dense ECM microstructure can result in increased fluid pressure within the device, which could result in cell damage and further influence drug responses and tumor resistance [57]. Moreover, a recent tendency in ECM modeling in vitro involves matrix stiffness mimicry [38,56], as it greatly contributes to the tumor phenotype [58]; therefore, we believe that successful tumor matrix mimicry would be a major step for tumor-on-chip development.…”

A Wireless Implantable Strain Sensing Scheme Using Ultrasound Imaging of Highly Stretchable Zinc Oxide/Poly Dimethylacrylamide Nanocomposite Hydrogel

“…The antibody against cell survival and apoptosis marker caspase-3 (Cell Signaling Technologies, Boston, MA; 0.129 μg/mL) was used in conjunction with the Texas red-conjugated secondary antibody (Life Technologies, Carlsbad, CA; 2 μg/mL) to test the toxicity of each of the substrate materials studied. Cells were fixed with 4% formaldehyde (Sigma-Aldrich) and processed for immunostaining, as described previously. , To visualize the cytoskeleton, actin filaments were stained with Alexa Fluor 488-conjugated phalloidin (Thermo Fisher; 125 μg/mL). Nuclei were counter-stained with 500 μg/mL 4′,6-diamidino-2-phenylindole (DAPI).…”

“…Cells were fixed with 4% formaldehyde (Sigma-Aldrich) and processed for immunostaining, as described previously. 47,48 To visualize the cytoskeleton, actin filaments were stained with Alexa Fluor 488-conjugated phalloidin (Thermo Fisher; 125 μg/mL). Nuclei were counter-stained with 500 μg/mL 4′,6-diamidino-2phenylindole (DAPI).…”

Mechanical Studies of the Third Dimension in Cancer: From 2D to 3D Model

“…The inclusion of mechanical constraints in designing an in vitro model requires the use of 3D culture platforms (scaffold-free or scaffold-based approaches) to fully mimic native tumor tissue biology as well as mechanical and biochemical properties [ 49 ]. The use of scaffold-based approaches to growing cells in a 3D environment is very common in tissue engineering [ 50 ].…”