TrypLE reagent product image

Gibco TrypLE gentle cell dissociation reagents

Ideal for both serum-supplemented and serum-free conditions, TrypLE enzymes are an answer to animal origin-free trypsin-like enzymes. Exhibiting a similar pH activity profile to trypsin, this enzyme cleaves at arginine and lysine. With the TrypLE enzyme’s animal-free origin, hazards from potential pathogenic contaminants have been eliminated; and subject to a controlled fermentation process, TrypLE enzymes are available for supply at any scale.

The TrypLE enzyme is designed to seamlessly fit your workflow. TrypLE enzymes can be substituted for trypsin in existing protocols, is room temperature stable and, in addition, is exceedingly gentle on cells. So gentle, in fact, that the TrypLE enzyme requires no inhibiting agent. Instead, it is inactivated by dilution alone. In addition, TrypLE enzymes boast similar dissociation kinetics to porcine trypsin as well as comparable cell replating, proliferation kinetics, and long-term maintenance.

TrypLE products offer a greener, more economical cell dissociation. For more information, see our TrypLE enzyme resources

Choose the right TrypLE enzymes for your cell culture

 TrypLE ExpressTrypLE Select
CharacteristicsGentler, stabler, and cost comparableGentler, stabler, animal and human-component free
Phenol redAvailable with or without phenol redPhenol red-free
Use caseGeneral purposeBioproduction/industrial application
Origin sourceFungus
(animal origin-free)
Fungus
(animal origin-free)
Manufacturing standardscGMP manufacturingcGMP, manufactured on dedicated animal origin-free machinery
Storage temperatureRoom temperatureRoom temperature
Higher cell viability
Inactivation methodInhibition by dilutionInhibition by dilution

A clear bottle on white background containing TrypLE Express Enzyme, with Phenol red

TrypLE Express reagents

Using the same recombinant fungal trypsin-like protease, the TrypLE Express reagent is manufactured to accommodate pricing comparable to standard trypsin. Unlike TrypLE Select formulations, TrypLE Express reagents come with and without phenol red.

For data on release times, stability statistics, and purity and plating efficiency, please see TrypLE Express performance data below.

TrypLE Express performance data

Cell line release times and viability

Triplicate cultures of each cell line, serially passed six times over, were centrifugally washed, counted, and subcultured post-enzyme-dissociation. The following values represent replicate averages over these six passages.

Cell lineMediaMean time required
for cell release
Mean viability
VEROVP-SFM5 min100%
VEROMEM + 5% FBS8 min100%
VEROOptiPro7 min98%
PK-15MEM + 10% FBS27 min98%
PK-15OptiPro11 min99%
MDCKOptiPro28 min98%
MDBKDMEM + 5% FBS15 min100%
A549DMEM + 10% FBS9 min98%
293FDMEM + 5% FBS2 min97%

TrypLE Express reagent outperforms trypsin in preserving cell-surface epitope expression

Histogram showing how TrypLE Express reagent preserves CD2-APC better than trypsin

Figure 1. TrypLE Express reagent preserves cell-surface epitope expression. Jurkat cells were treated with 0.05% trypsin (A) or TrypLE Express reagent (B) for a period of up to 20 minutes. Cell-surface CD2 levels were then measured via flow cytometry with an APC-conjugated anti-CD2 monoclonal antibody. As demonstrated, cells treated with 0.05% trypsin show a clear time-dependent reduction in CD2 levels, those treated with TrypLE Express reagent exhibit no measurable CD2 loss.

Flow cytometry analysis: cell dissociation from mouse CNS tissue

Bar chart of viability vs annexin V when using TrypLE enzymes or Accutase

Figure 2. Cell viability is better with TrypLE reagent than with Accutase reagent​.
Dissociation time: 10 min; Viability (%) +/- SEM​. Adapted from data shown in Panchision DM, et al. Stem Cells, 25:1560 (2007). doi:10.1634/stemcells.2006-0260.

Bar chart of percent CD24-positive cells for both TrypLE reagent and papain treatments

Figure 3. The CD24 epitope is retained with TrypLE whereas use of papain illustrates complete loss of the CD24 epitope. Adapted from data shown in Panchision DM, et al. Stem Cells, 25:1560 (2007). doi:10.1634/stemcells.2006-0260.

Workflow benefits when using TrypLE enzymes

TrypLE enzymes simplify your workflow. For example, cells, treated with either animal trypsin or TrypLE Express, and replated with OptiPro SFM were left unwashed post-dissociation. In addition, no protease inhibitors were used. 24 hours later, morphology is recorded. As seen below, TrypLE enzyme-treated cells retain their structure, while trypsin cells experience further cellular degradation. 

Animal trypsin
Brightfield image of cultured cells after trypsin treatment
TrypLE Express
Brightfield image of cultured cells after TrypLE enzyme treatment

Figure 4. TrypLE enzyme-treated cells retain their structure when left unwashed after dissociation without protease inhibitors. While TrypLE enzyme-treated cells maintained their morphology under these conditions, trypsin-treated cells experienced further degradation.

Cell line release times and viability

Triplicate cultures of each cell line, serially passed six times over, were centrifugally washed, counted, and subcultured post-enzyme-dissociation. The following values represent replicate averages over these six passages.

Cell lineMediaMean time required
for cell release
Mean viability
VEROVP-SFM5 min100%
VEROMEM + 5% FBS8 min100%
VEROOptiPro7 min98%
PK-15MEM + 10% FBS27 min98%
PK-15OptiPro11 min99%
MDCKOptiPro28 min98%
MDBKDMEM + 5% FBS15 min100%
A549DMEM + 10% FBS9 min98%
293FDMEM + 5% FBS2 min97%

TrypLE Express reagent outperforms trypsin in preserving cell-surface epitope expression

Histogram showing how TrypLE Express reagent preserves CD2-APC better than trypsin

Figure 1. TrypLE Express reagent preserves cell-surface epitope expression. Jurkat cells were treated with 0.05% trypsin (A) or TrypLE Express reagent (B) for a period of up to 20 minutes. Cell-surface CD2 levels were then measured via flow cytometry with an APC-conjugated anti-CD2 monoclonal antibody. As demonstrated, cells treated with 0.05% trypsin show a clear time-dependent reduction in CD2 levels, those treated with TrypLE Express reagent exhibit no measurable CD2 loss.

Flow cytometry analysis: cell dissociation from mouse CNS tissue

Bar chart of viability vs annexin V when using TrypLE enzymes or Accutase

Figure 2. Cell viability is better with TrypLE reagent than with Accutase reagent​.
Dissociation time: 10 min; Viability (%) +/- SEM​. Adapted from data shown in Panchision DM, et al. Stem Cells, 25:1560 (2007). doi:10.1634/stemcells.2006-0260.

Bar chart of percent CD24-positive cells for both TrypLE reagent and papain treatments

Figure 3. The CD24 epitope is retained with TrypLE whereas use of papain illustrates complete loss of the CD24 epitope. Adapted from data shown in Panchision DM, et al. Stem Cells, 25:1560 (2007). doi:10.1634/stemcells.2006-0260.

Workflow benefits when using TrypLE enzymes

TrypLE enzymes simplify your workflow. For example, cells, treated with either animal trypsin or TrypLE Express, and replated with OptiPro SFM were left unwashed post-dissociation. In addition, no protease inhibitors were used. 24 hours later, morphology is recorded. As seen below, TrypLE enzyme-treated cells retain their structure, while trypsin cells experience further cellular degradation. 

Animal trypsin
Brightfield image of cultured cells after trypsin treatment
TrypLE Express
Brightfield image of cultured cells after TrypLE enzyme treatment

Figure 4. TrypLE enzyme-treated cells retain their structure when left unwashed after dissociation without protease inhibitors. While TrypLE enzyme-treated cells maintained their morphology under these conditions, trypsin-treated cells experienced further degradation.


A clear bottle with a white label on a white background containing TrypLE Select Enzyme (1X), Phenol red free

TrypLE Select reagents

TrypLE Select enzymes are manufactured on dedicated animal origin-free machinery and designed specifically for the bioproduction industry. Select enzymes come in two concentrations, 1X and 10X, and are phenol red-free. Using 10X at full strength can rapidly, but gently, dissociate strongly adherent cells.

For data on release times, stability statistics, and purity and plating efficiency, please see TrypLE Select data below.

TrypLE Select performance data

Cell release times

The following cell lines were treated with TrypLE Select enzymes at either 1X or 10X concentrations. Their mean cell release times and mean viabilities were observed and recorded.

 (1x)(10x)
StrengthCell linesMediaMean cell
release time
Mean
viability
Mean cell
release time
Mean
viability
Adherent293FDMEM + 5% FBS2 min97%  
VEROVP-SFM5 min100%  
VEROEMEM + 5% FBS8 min100%  
VEROOptiPRO SFM7 min98%  
CHO-K1CHO III A7 min95%  
Strongly adherentPK-15EMEM + 10% FBS27 min98%11.7±2.0 min99.4%±0.3%
PK-15Gibco OptiPRO SFM11 min99%  
MDCKOptiPRO SFM28 min98%15.3±0.8 min98.7%±0.5%
MDCKDMEM + 5% FBS15 min100%8.0±1.3 min98.8%±0.5%
A549DMEM + 10% FBS9 min98%5.2±0.3 min99.1%±0.5%
MSCDMEM + 10% MSC FBS  7.8±1.2 min98.7%±1.3%
MSCGibco StemPro MSC SFM  8.7±1.5 min97.5%±0.1%

Stability statistics

At various temperatures, the TrypLE Select enzyme shows superior stability compared to either crude or purified porcine trypsin. Even after eight weeks at 37°C, the TrypLE Select enzyme maintained 50% activity while, after only two days, purified porcine trypsin enzyme activity rapidly decreased. 

Bar chart of TrypLE Select enzyme activity over 8 weeks

Figure 5. TrypLE Select enzyme stability. TrypLE Select enzyme shows superior stability compared to crude or purified porcine trypsin at various storage temperatures. Even after eight weeks at 37°C, 50% enzyme activity was still seen. All samples were stored in the dark.

Bar chart of porcine trypsin activity over 8 weeks
Figure 6. Purified porcine trypsin stability. Enzyme activity decreased rapidly in samples of purified porcine trypsin stored at 37°C.

Plating efficiency

Cells detached with TrypLE Select enzymes at 100–200 cells per well, have higher plating efficiency than those treated with porcine trypsin. 

Bar chart of percent animal enzyme control for 2 cell lines and TrypLE Select enzyme and animal trypsin treatments
Figure 7. TrypLE Select enzymes increases plating efficiency. Cells detached with TrypLE Select enzymes exhibit higher plating efficiency than cells detached with porcine trypsin.

Cell release times

The following cell lines were treated with TrypLE Select enzymes at either 1X or 10X concentrations. Their mean cell release times and mean viabilities were observed and recorded.

 (1x)(10x)
StrengthCell linesMediaMean cell
release time
Mean
viability
Mean cell
release time
Mean
viability
Adherent293FDMEM + 5% FBS2 min97%  
VEROVP-SFM5 min100%  
VEROEMEM + 5% FBS8 min100%  
VEROOptiPRO SFM7 min98%  
CHO-K1CHO III A7 min95%  
Strongly adherentPK-15EMEM + 10% FBS27 min98%11.7±2.0 min99.4%±0.3%
PK-15Gibco OptiPRO SFM11 min99%  
MDCKOptiPRO SFM28 min98%15.3±0.8 min98.7%±0.5%
MDCKDMEM + 5% FBS15 min100%8.0±1.3 min98.8%±0.5%
A549DMEM + 10% FBS9 min98%5.2±0.3 min99.1%±0.5%
MSCDMEM + 10% MSC FBS  7.8±1.2 min98.7%±1.3%
MSCGibco StemPro MSC SFM  8.7±1.5 min97.5%±0.1%

Stability statistics

At various temperatures, the TrypLE Select enzyme shows superior stability compared to either crude or purified porcine trypsin. Even after eight weeks at 37°C, the TrypLE Select enzyme maintained 50% activity while, after only two days, purified porcine trypsin enzyme activity rapidly decreased. 

Bar chart of TrypLE Select enzyme activity over 8 weeks

Figure 5. TrypLE Select enzyme stability. TrypLE Select enzyme shows superior stability compared to crude or purified porcine trypsin at various storage temperatures. Even after eight weeks at 37°C, 50% enzyme activity was still seen. All samples were stored in the dark.

Bar chart of porcine trypsin activity over 8 weeks
Figure 6. Purified porcine trypsin stability. Enzyme activity decreased rapidly in samples of purified porcine trypsin stored at 37°C.

Plating efficiency

Cells detached with TrypLE Select enzymes at 100–200 cells per well, have higher plating efficiency than those treated with porcine trypsin. 

Bar chart of percent animal enzyme control for 2 cell lines and TrypLE Select enzyme and animal trypsin treatments
Figure 7. TrypLE Select enzymes increases plating efficiency. Cells detached with TrypLE Select enzymes exhibit higher plating efficiency than cells detached with porcine trypsin.

TrypLE enzyme resources

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