Trypsin Medium
Trypsin medium refers to a solution containing the proteolytic enzyme trypsin, which is used for detaching adherent cells from culture surfaces during subculturing and harvesting. It is particularly important in cell culture workflows to disaggregate cells for reseeding, analysis, or storage. Trypsin cleaves peptide bonds at the carboxyl side of lysine and arginine residues, efficiently breaking cell-cell and cell-matrix interactions.
Content
The typical composition of trypsin medium includes:
- Trypsin Enzyme
- A protease (often porcine-derived) at concentrations ranging from 0.05% to 0.25%, depending on the application and cell sensitivity.
- EDTA (Ethylenediaminetetraacetic Acid)
- Added at 0.02%–0.05% to chelate divalent cations (e.g., Ca²⁺ and Mg²⁺) and enhance trypsin activity by disrupting cadherin-mediated cell adhesion.
- Buffer System
- Typically buffered with phosphate-buffered saline (PBS) or HEPES to maintain physiological pH (7.2–7.4).
- Salts
- Sodium chloride, potassium chloride, and other salts to maintain osmotic balance.
Applications
- Cell Detachment for Subculturing
- Trypsin medium is used to detach adherent cells from culture flasks or plates for passaging, reseeding, or expanding cultures.
- Tissue Dissociation
- Facilitates the enzymatic digestion of tissues to obtain single-cell suspensions, often combined with other enzymes like collagenase for this purpose.
- Flow Cytometry and Sorting
- Used to prepare single-cell suspensions for fluorescence-activated cell sorting (FACS) and analysis.
- Cryopreservation Preparation
- Cells are detached using trypsin prior to freezing and storage in cryoprotective media.
Trypsin medium is an essential tool in cell culture workflows for detaching adherent cells and preparing single-cell suspensions. Its effectiveness depends on appropriate concentration, incubation time, and neutralization. Understanding the properties and handling requirements of trypsin medium ensures optimal cell viability and reproducibility in experimental workflows.
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