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TrypLE Enzymes
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 Express | TrypLE Select | |
|---|---|---|
| Characteristics | Gentler, stabler, and cost comparable | Gentler, stabler, animal and human-component free |
| Phenol red | Available with or without phenol red | Phenol red-free |
| Use case | General purpose | Bioproduction/industrial application |
| Origin source | Fungus (animal origin-free) | Fungus (animal origin-free) |
| Manufacturing standards | cGMP manufacturing | cGMP, manufactured on dedicated animal origin-free machinery |
| Storage temperature | Room temperature | Room temperature |
| Higher cell viability | ✓ | ✓ |
| Inactivation method | Inhibition by dilution | Inhibition by dilution |
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.
TrypLE Express reagent outperforms trypsin in preserving cell-surface epitope expression
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
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.
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
TrypLE Express
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.
TrypLE Express reagent outperforms trypsin in preserving cell-surface epitope expression
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
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.
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
TrypLE Express
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.
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) | |||||
|---|---|---|---|---|---|---|
| Strength | Cell lines | Media | Mean cell release time | Mean viability | Mean cell release time | Mean viability |
| Adherent | 293F | DMEM + 5% FBS | 2 min | 97% | ||
| VERO | VP-SFM | 5 min | 100% | |||
| VERO | EMEM + 5% FBS | 8 min | 100% | |||
| VERO | OptiPRO SFM | 7 min | 98% | |||
| CHO-K1 | CHO III A | 7 min | 95% | |||
| Strongly adherent | PK-15 | EMEM + 10% FBS | 27 min | 98% | 11.7±2.0 min | 99.4%±0.3% |
| PK-15 | Gibco OptiPRO SFM | 11 min | 99% | |||
| MDCK | OptiPRO SFM | 28 min | 98% | 15.3±0.8 min | 98.7%±0.5% | |
| MDCK | DMEM + 5% FBS | 15 min | 100% | 8.0±1.3 min | 98.8%±0.5% | |
| A549 | DMEM + 10% FBS | 9 min | 98% | 5.2±0.3 min | 99.1%±0.5% | |
| MSC | DMEM + 10% MSC FBS | 7.8±1.2 min | 98.7%±1.3% | |||
| MSC | Gibco StemPro MSC SFM | 8.7±1.5 min | 97.5%±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.
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.
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.
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) | |||||
|---|---|---|---|---|---|---|
| Strength | Cell lines | Media | Mean cell release time | Mean viability | Mean cell release time | Mean viability |
| Adherent | 293F | DMEM + 5% FBS | 2 min | 97% | ||
| VERO | VP-SFM | 5 min | 100% | |||
| VERO | EMEM + 5% FBS | 8 min | 100% | |||
| VERO | OptiPRO SFM | 7 min | 98% | |||
| CHO-K1 | CHO III A | 7 min | 95% | |||
| Strongly adherent | PK-15 | EMEM + 10% FBS | 27 min | 98% | 11.7±2.0 min | 99.4%±0.3% |
| PK-15 | Gibco OptiPRO SFM | 11 min | 99% | |||
| MDCK | OptiPRO SFM | 28 min | 98% | 15.3±0.8 min | 98.7%±0.5% | |
| MDCK | DMEM + 5% FBS | 15 min | 100% | 8.0±1.3 min | 98.8%±0.5% | |
| A549 | DMEM + 10% FBS | 9 min | 98% | 5.2±0.3 min | 99.1%±0.5% | |
| MSC | DMEM + 10% MSC FBS | 7.8±1.2 min | 98.7%±1.3% | |||
| MSC | Gibco StemPro MSC SFM | 8.7±1.5 min | 97.5%±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.
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.
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.
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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