Product image of trypsin

Detach adherent cells and tissues with Gibco cell dissociation reagents

To acquire a suitable number of cells for research, a group of cells must first be disaggregated from a larger culture. Enter: cell dissociation. Gibco trypsin and alternative cell dissociation products are ideal for use with tissues and cell monolayers. Available in a variety of formats, both enzymatic and chemical, to meet the varied needs of researchers performing adherent cell culture. Explore the selection guide below to choose the correct cell dissociation reagent for your cell culture type.

Explore dissociation protocols

Choose the best cell dissociation reagent for your cell culture type

Cell culture typeWash solutionDissociation methodRecommended productFormatContains phenol red
Most continuous and strongly adherent cell linesHBSS or PBS w/o calcium and magnesiumTrypsin 0.25% in balanced salt solution w/o calcium and magnesium0.25% Trypsin (1X)LiquidYes
2.5% Trypsin (10X)LiquidNo
Trypsin (1:250)PowderNo
Many cells at early passageD-PBSTrypLE reagentTrypLE ExpressLiquidYes
Continuous cell lines where cell surface protein integrity is importantHBSS or PBS w/o calcium and magnesiumTrypsin 0.05% in 0.53 mM EDTA0.05% Trypsin 0.53 mM EDTALiquidYes
0.05% Trypsin 0.53 mM EDTA (10X)LiquidNo
TrypLE reagentTrypLE ExpressLiquidYes
Weakly adherent epithelial cellsHBSS or PBS w/o calcium and magnesiumEDTA, glycerol in sodium citrateCell Dissociation Buffer, Hanks'-basedLiquidNo
Cell Dissociation Buffer, PBS-basedLiquidNo
Transformed fibroblasts
HBSS or PBS w/o calcium and magnesiumTrypLE reagentTrypLE Express (1X)LiquidYes
Primary cells where cell surface protein integrity is importantHBSS or PBS w/o calcium and magnesiumTrypLE reagentTrypLE Express (1X)LiquidYes
Strongly adherent early-passage cell linesHBSS or PBS w/o calcium and magnesiumTrypsin 0.25% in 1 mM EDTA0.25% Trypsin 1 mM EDTALiquidYes
Trypsin Powder (1:250) PowderNo
Dispase 0.6–2.4 units/mL in PBSDispase IIPowder, LyophilizedNo
Epithelial cells0.5–1 mM EDTA0.5–1 mM EDTAVersene solution 1:5,000 (0.53 mM EDTA in PBS)LiquidNo
Dispase 0.6–2.4 units/mL in PBSDispase IIPowder, LyophilizedNo
Strongly adherent, epithelial and some tumor cell lines0.5–1 mM EDTA0.25% with 1 mM EDTA0.25% Trypsin 1 mM EDTALiquidYes
Versene solution 1:5,000 (0.53 mM EDTA in PBS)LiquidNo
Trypsin Powder (1:250) PowderNo
Dispase 0.6–2.4 units/mL in PBS w/o calcium and magnesiumDispase IIPowder, LyophilizedNo
Thick cultures, multiple layers1 mM EDTATrypsin 0.25%0.25% Trypsin 1 mM EDTALiquidYes
Trypsin Powder (1:250)PowderNo
Dense cultures that are collagen rich1 mM EDTACollagenase 200 units/mL, 1 mM EDTA in balanced salt solution w/o calcium and magnesiumCollagenase Type IPowder, LyophilizedNo
Collagenase Type IIPowder, LyophilizedNo
Collagenase Type IVPowder, LyophilizedNo
All adherent cell culturesHBSS or PBS w/o calcium and magnesiumPhysical/mechanicalCell scrapers

Trypsin and cell trypsinization

Trypsin, a proteolytic enzyme, is the standard way to detach adherent cell cultures and monolayers. This globular, pancreatic protease cleaves at the C-terminal side of lysine and arginine, breaking down vessel-adhering proteins and allowing easy resuspension during cell harvesting.

Thermo Fisher Scientific offers porcine-derived trypsin, EDTA-enhanced trypsin, as well as formulations of the TrypLE enzyme, a gentle, trypsin-like enzyme. To avoid unplanned protein degradation, organic trypsin must be inactivated with trypsin inhibitors. In contrast, synthetic trypsin, such as Gibco TrypLE products, are deactivated via dilution, eliminating the need for additional solutions.

NOTE: If you have fetal bovine serum (FBS) in your cell culture media, the culture will need to be washed with a balanced salt solution before trypsinization to ensure trypsin functionality.

Product image of trypsin
Trypsin-EDTA (0.25%), phenol red: a medium clear bottle with white and green labeling

Trypsin EDTA  

EDTA, a chelating agent, improves trypsin’s ability to detach adherent cells. Binding calcium and magnesium into a hexadentate, EDTA weakens cell-cell adhesion and enhances trypsin’s access to hydrolysis-targeted peptide bonds.

Gibco Trypsin-EDTA is made from irradiated porcine-derived protease with e-beam verification as well as tested for PPV, mycoplasma, and PCV 1/2 contamination.


 TrypsinTrypsin-EDTATrypLE ExpressTrypLE Select
CharacteristicsTraditional standard
Trypsin with enhanced cleavageGentler, stabler, and cost comparableGentler, stabler, animal and human-component free
Use caseGeneral purposeGeneral purposeGeneral purposeBioproduction/industrial application
Origin sourcePorcine pancreasPorcine pancreasFungus (animal origin-free)Fungus (animal origin-free)
Manufacturing standardscGMP manufacturingcGMP manufacturingcGMP manufacturingcGMP, manufactured on dedicated animal origin-free machinery
Storage temperature–5 to –20° C–5 to –20° CRoom temperatureRoom temperature
Higher cell viability  
Inactivation methodTrypsin inhibitorsTrypsin inhibitorsInhibition by dilutionInhibition by dilution

Trypsin inhibitors

Trypsin inhibitors destructively alter trypsin, nullifying the proteolytic enzyme’s ability to bind with, and so digest, proteins. Used post dissociation, trypsin inhibitors protect your cells against further protein degradation by trypsin.

NOTE: Soybean trypsin inhibitors also bind chymotrypsin, albeit to a lesser degree.

Trypsin inhibitors comparison chart

 Soybean trypsin inhibitor (powder)Defined trypsin inhibitorTrypsin neutralizer solution
Animal origin-free  
Calcium and magnesium free  
With 0.5% newborn bovine serum  


Relatively gentle, collagenase works by digesting connective tissue’s native triple-helical collagen fibrils. These tissues are often found in skin, tendon, blood vessels, and bone—making collagenase disaggregation suitable for human tumor, mouse kidney, human brain, and epithelium cultures.

Visit the Technical Reference Library to view protocols for collagenase dissociation of cells from primary tissue

Choose the right collagenase for your cell type

CollagenaseTissue/Cell type
Type IEpithelial, adrenal, lung, fat
Type IIHeart, thyroid, salivary, liver, bone, cartilage
Type IVIslet (insulin receptor sites)


Dispase, or neutral protease, is a Bacillus polymyxa–produced metalloenzyme. Rapid and gentle, dispase can be used to harvest and transfer normal diploid cells and cell lines. Dispase effectively separates cell clumps and splits cells from intact tissue with little impact to membrane integrity or viability.

Non-enzymatic cell dissociation

Versene solution

Versene solution, an EDTA solution in PBS, dissociates cells gently and non-enzymatically. This chelating agent detaches some cell types (such as epithelial cells) from vessel surfaces for suspension. It can also be used as a wash prior to trypsinization.

For further guidance on Versene solution applications, please see the cell dissociation selection guide by culture type.

Non-enzymatic cell dissociation buffers

Recommended for weakly adherent cells, such as epithelial cells, Gibco cell dissociation buffers dissociate gently and maintain surface protein integrity. Applications include cells involved in ligand binding, flow cytometry, and immunohistochemistry research.

Please consult the enzyme-free cell dissociation buffer protocol for more information.

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Gibco cell culture products are manufactured in facilities compliant with current good manufacturing practices (GMP) and adhere to a robust quality management system, helping to ensure the consistency, reliability, and high quality you can rely on.

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