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Tryple cell dissociation reagent

Manufactured by Thermo Fisher Scientific
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Sourced in United States, United Kingdom

TrypLE is a cell dissociation reagent used to detach adherent cells from cell culture surfaces. It is a recombinant trypsin-like protease that cleaves the peptide bonds in cell surface proteins, allowing cells to be dissociated from the substrate. TrypLE is a ready-to-use, animal-origin-free alternative to traditional trypsin-based cell dissociation reagents.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Y 27632 dihydrochloride, by Bio-Techne (3 mentions) Human bfgf, by Bio-Techne (3 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (3 mentions) Dmem f12, by Thermo Fisher Scientific (3 mentions) β mercaptoethanol, by Merck Group (3 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions) Facs tubes, by Greiner (1 mentions) Sytox red, by Thermo Fisher Scientific (1 mentions) Canto 2, by BD (1 mentions) Cd13 bv421, by BioLegend (1 mentions) Macsquant analyzer, by Miltenyi Biotec (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) 96 well plate, by Corning (1 mentions)

Close spelling variants of this product

TrypLE express cell dissociation reagent TrypLE dissociation reagent Cell Dissociation Reagent TrypLE reagent TrypLE

6 protocols using tryple cell dissociation reagent

1

Maintaining and Differentiating Human iPSCs

Cited 2 times
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The iPSCs were maintained on irradiated Mouse Embryonic Fibroblasts (MEFs) in 6-well tissue culture plates at 37 °C with 5% CO2 and 5% O2 They were grown in human embryonic stem cell (hESC-10) medium made up of a base of DMEM/F12 (50:50 Gibco) with 20% knock-out serum replacement (KOSR, Gibco), 100 μM nonessential amino acids, 2 mM glutamine, 50 U/ml penicillin, 50 μg/ml streptomycin (all from Invitrogen), 10−4 M β–mercaptoethanol (Sigma, St. Louis, MO), and 10 ng/ml human bFGF (BioTechne). Cells were passaged every 5–7 days once they reached about 70–80% confluency using TrypLE cell dissociation reagent (Gibco) and gentle scraping before being replated onto new MEFs. Cells were transitioned in hESC-10 medium with 10 μM ROCK inhibitor (Y27632 dihydrochloride, Tocris) for <24 h before replacing the media with fresh hESC-10 medium without ROCK inhibitor. Directed differentiations were performed essentially as described for mesoderm (Thom et al., 2020 (link)), endoderm (Leavens et al., 2021 (link)), and ectoderm (Telezhkin et al., 2016 (link)).
Wilken M.B., Maguire J.A., Dungan L.V., Gagne A., Osorio-Quintero C., Waxman E.A., Chou S.T., Gadue P., French D.L, & Thom C.S. (2023). Generation of a human Tropomyosin 1 knockout iPSC line. Stem cell research, 71, 103161.
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2

Maintaining and Differentiating Human iPSCs

Cited 2 times
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The iPSCs were maintained on irradiated Mouse Embryonic Fibroblasts (MEFs) in 6-well tissue culture plates at 37 °C with 5% CO2 and 5% O2 They were grown in human embryonic stem cell (hESC-10) medium made up of a base of DMEM/F12 (50:50 Gibco) with 20% knock-out serum replacement (KOSR, Gibco), 100 μM nonessential amino acids, 2 mM glutamine, 50 U/ml penicillin, 50 μg/ml streptomycin (all from Invitrogen), 10−4 M β–mercaptoethanol (Sigma, St. Louis, MO), and 10 ng/ml human bFGF (BioTechne). Cells were passaged every 5–7 days once they reached about 70–80% confluency using TrypLE cell dissociation reagent (Gibco) and gentle scraping before being replated onto new MEFs. Cells were transitioned in hESC-10 medium with 10 μM ROCK inhibitor (Y27632 dihydrochloride, Tocris) for <24 h before replacing the media with fresh hESC-10 medium without ROCK inhibitor. Directed differentiations were performed essentially as described for mesoderm (Thom et al., 2020 (link)), endoderm (Leavens et al., 2021 (link)), and ectoderm (Telezhkin et al., 2016 (link)).
Wilken M.B., Maguire J.A., Dungan L.V., Gagne A., Osorio-Quintero C., Waxman E.A., Chou S.T., Gadue P., French D.L, & Thom C.S. (2023). Generation of a human Tropomyosin 1 knockout iPSC line. Stem cell research, 71, 103161.
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3

Culturing and Directed Differentiation of iPSCs

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The iPSCs were maintained on irradiated Mouse Embryonic Fibroblasts (MEFs) in 6-well tissue culture plates at 37°C with 5% CO2 and 5% O2 They were grown in human embryonic stem cell (hESC-10) medium made up of a base of DMEM/F12 (50:50 Gibco) with 20% knock-out serum replacement (KOSR, Gibco), 100 μM nonessential amino acids, 2 mM glutamine, 50 U/ml penicillin, 50 μg/ml streptomycin (all from Invitrogen), 10−4 M β–mercaptoethanol (Sigma, St. Louis, MO), and 10 ng/ml human bFGF (BioTechne). Cells were passaged every 5–7 days once they reached about 70–80% confluency using TrypLE cell dissociation reagent (Gibco) and gentle scraping before being replated onto new MEFs. Cells were transitioned in hESC-10 medium containing with 10 μM ROCK inhibitor (Y27632 dihydrochloride, Tocris) for <24 hours before replacing the media with fresh hESC-10 medium without ROCK inhibitor. Directed differentiations were performed essentially as described for mesoderm (Thom et al., 2020 ), endoderm (Leavens et al., 2021 ), and ectoderm (Telezhkin et al., 2016 ).
Wilken M.B., Maguire J.A., Dungan L.V., Gagne A., Osorio-Quintero C., Waxman E.A., Chou S.T., Gadue P., French D.L, & Thom C.S. (2023). Generation of a human Tropomyosin 1 knockout iPSC line. bioRxiv.
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4

Cytotoxicity and Phagocytosis Assay

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At the end of the co-culture, supernatants were collected into FACS tubes (Greiner Bio-One, North Carolina, USA), and cells were washed with PBS once. As the manufacturer suggested, TrypLE cell dissociation reagent (Gibco, ThermoFisher Scientific, Waltham, MA, USA) was used to detach cells. Then, cells were collected, transferred into FACS tubes, and washed twice with PBS. At the last step, they were stained with 5 nM SYTOX Red in PBS containing 0.5% BSA (ThermoFisher Scientific, Waltham, MA, USA) for 15 minutes on ice in the dark. Subsequently, both the cytotoxicity of macrophages for AML cells (Tag-it Violet+ CFSE- SYTOX Red+) and the phagocytosis of AML cells by live macrophages (CFSE+ SYTOX Red- Tag-it Violet+) were assessed by Canto II (BD Biosciences). The data were analyzed with the FlowJo software (TreeStar, Ashland, OR). Macrophage cytotoxicity for tumor cells was calculated as % of SYTOX Red+ AML cells (Tag-it Violet+ CFSE-) co-cultured with macrophages (CFSE+)- % SYTOX Red+ AML cells alone.
Gunalp S., Helvaci D.G., Oner A., Bursalı A., Conforte A., Güner H., Karakülah G., Szegezdi E, & Sag D. (2023). TRAIL promotes the polarization of human macrophages toward a proinflammatory M1 phenotype and is associated with increased survival in cancer patients with high tumor macrophage content. Frontiers in Immunology, 14, 1209249.
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5

Integrin Expression in APAP Hepatotoxicity

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Integrin expression in response to APAP. To assess correlation of z-alpha with modulation of expression of integrins, adhesion molecules and other markers in response to APAP hepatotoxicity, remaining adherent HepaRG cells were recovered using TrypLE™ cell-dissociation reagent (Life Technologies), and stained using combinations of directly conjugated monoclonal antibodies: CD29-BV510 (563513), CD49f-BV421 (5625820, CD49d-FITC (580840), CD49c-PE, CD166-PE (559263) (all BD Biosciences, Oxford, UK); CD49a-APC-Vio770 (130-101-406), CD44-APC-Vio770 (130-099-149), CD49b-PE-Vio770 (130-100-328), CD90-PE-Vio770 (130-099-295), CD49e-APC (130-097-221), (all Miltenyi Biotec, Surrey, UK); CD13-BV421 (301716; Biolegend), CD54-FITC (mhcd5401; Caltag, Buckingham, UK). Cells were incubated with optimal concentration of antibodies (1/50) at 4 °C for 20 minutes, washed twice and resuspended in PBS containing 0.1% BSA and 0.1% sodium azide. Unstained cells were included as controls, and dead cells and debris were excluded from the analysis, based on scatter characteristics. Data for at least 10,000 live events per sample were acquired using a MACSQuant Analyzer (Miltenyi Biotec) and analyzed using FlowJo version 9.6.7 software (Flowjo LLC). Data is expressed as relative mean fluorescence intensity (MFI), and % positive staining.
Gamal W., Treskes P., Samuel K., Sullivan G.J., Siller R., Srsen V., Morgan K., Bryans A., Kozlowska A., Koulovasilopoulos A., Underwood I., Smith S., del-Pozo J., Moss S., Thompson A.I., Henderson N.C., Hayes P.C., Plevris J.N., Bagnaninchi P.O, & Nelson L.J. (2017). Low-dose acetaminophen induces early disruption of cell-cell tight junctions in human hepatic cells and mouse liver. Scientific Reports, 7, 37541.
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6

Nanomaterial Uptake in Cell Lines

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For adherent cell lines (HeLa and NIH/3T3), nanotubes were introduced to a monolayer of cells at a concentration of 5 mg/L for 5 min in cell media without serum. After this time, they were washed 3× with PBS (Gibco), and placed in fresh cellular media without serum for data acquisition. In the case of the suspension cell line, Jurkat, 5 mg/L nanotubes were added to a microcentrifuge tube containing 106 cells. The solution was gently shaken for 5 min, followed by 2 min of centrifugation at 3000g. The cells were resuspended with a pipet tip in cell media without serum, and then centrifuged again. This process was repeated 3× in order to remove all unbound nanotubes from the solution. A volume of 100 uL in a 96-well plate (Corning) was used for excitation/ emission spectroscopy.
For trypsinization experiments, 106 cells were incubated with TrypLE cell-dissociation reagent (Life technologies), a recombinant porcine trypsin alternative, for 15 min to ensure all cells were detached from the substrate. The suspension was centrifuged at 3000g for 2 min, followed by resuspension with a pipet. This process was repeated 3× in order to remove the remaining enzyme. The cells were then incubated with nanotubes at 5 mg/L and washed in a similar fashion as the Jurkat procedure mentioned above.
Roxbury D., Jena P.V., Shamay Y., Horoszko C.P, & Heller D.A. (2015). Cell Membrane Proteins Modulate the Carbon Nanotube Optical Bandgap via Surface Charge Accumulation. ACS nano, 10(1), 499-506.
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