Three dimensional (3D) cell cultures are artificially-created environments in which cells are permitted to grow or interact with their surroundings in a 3D fashion. 3D cell cultures improve the function, differentiation and viability of cells and recapitulate in vivo microenvironment compared to conventional 2D cell cultures. 3D matrices provide a physiologically relevant screening platform, by mimicking the in vivo responses, for many cell types including cancer and stem cells in developmental morphogenesis, pharmacology, drug metabolism and drug toxicity studies. However, when it comes to cell passaging, or studying biochemical processes and protein analysis, there are challenges with separating intact cells from extra-cellular proteins comprising the 3D matrices. While proteases (such as trypsin and accutase) are commonly used to degrade these matrices, it is not recommended in some cases where cells are sensitive to protease digestion; these proteases may modify cellular proteins and signaling or cell surface, thus altering physiological assessments. Further, some proteases-based dissociation methods do not completely dissolve the matrices. Assay Genie's 3D Culture Non-Enzymatic Cell Harvesting Kit provides an optimized and standardized saline-based solution for the isolation of cells and spheroids from matrices (especially for Assay Genie's 3D Cell Culture Kits with high viability rate for subsequent biochemical, protein and cell-based analysis.

Figure: Dissociation of HepG2 cells from 3D cell culture Alginate Hydrogel or BME Matrix: Cells were cultured and formed spheroids in Alginate Hydrogel or BME Matrices for 21 days. Cells were then dissociated from matrices and harvested according to protocol above. Analysis using Calcein AM staining (green, for live cells), and Ethidium Homodimer-1 dye (red, for apoptotic cells) suggested that cell viability is not affected after performing harvesting protocol. Note: Calcein AM and Ethidium Homodimer-1 are not included in the kit.