This section provides guidelines and general procedures for routine subculturing, thawing, and freezing of cells in culture.
Note that cell culture conditions vary for each cell type. The consequences of deviating from the culture conditions required for a particular cell type can range from the expression of aberrant phenotypes to a complete failure of the cell culture. We therefore recommend that you familiarize yourself with your cell line of interest, and closely follow the instructions provided with each product you are using in your experiments.
Methods & Cell Culture Protocols
- Concentrating Cells: A procedure to concentrate cells from suspension culture or to resuspend cells from a monolayer culture.
- Counting Cells in a Hemocytometer: How to count and calculate the number of cells from a stock flask or culture dish.
- Counting Cells in a Countess II: How to count and calculate the number of cells using an automated cell counter.
- Counting Cells in a Countess II following TrypLE reagent cell dissociation: How to count cells using TrypLE reagent and an automated cell counter.
- 3D Cell Culture Protocols: Suggested protocols for generation of spheroids from the most commonly used cancer cell lines using Nunclon Sphera cell culture plastics and Gibco Media.
- Guidelines for Maintaining Cultured Cells: Learn what is subculture and when to do it, review media recommendations for common cell lines and how to dissociate cells as well as the benefits of using Gibco TrypLE as an alternative to trypsin.
- Subculturing Adherent Cells: A general procedure for subculturing adherent mammalian cells in culture.
- Subculturing Suspension Cells: A general procedure for subculturing mammalian and insect cells in suspension culture.
- Freezing Cells: A general procedure to properly cryopreserving cell lines.
- Thawing Frozen Cells: A general procedure for properly thawing cell lines.
- Trypan Blue Exclusion: A rapid assay to accurately determine the viability of your cells.
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