Trypsin

Manufactured by Lonza

Sourced in Belgium, Switzerland, United States, United Kingdom, Germany

Trypsin is a proteolytic enzyme used in cell culture applications to facilitate the detachment of adherent cells from the culture surface. It catalyzes the hydrolysis of peptide bonds, particularly those involving the carboxyl group of arginine or lysine.

Automatically generated - may contain errors

Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (24 mentions) Penicillin streptomycin, by Lonza (18 mentions) Ethylene diamine tetraacetic acid (edta), by Lonza (16 mentions) Phosphate buffered saline (pbs), by Lonza (14 mentions) Penicillin, by Lonza (11 mentions) Streptomycin, by Lonza (9 mentions) L glutamine, by Lonza (9 mentions) Fetal bovine serum (fbs), by Lonza (9 mentions) Dulbecco s modified eagle s medium dmem, by Lonza (8 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions) Dulbecco modified eagle medium (dmem), by Lonza (6 mentions) Fetal bovine serum (fbs), by Biowest (6 mentions) Fetal bovine serum (fbs), by Merck Group (5 mentions) Trypan blue, by Lonza (5 mentions) Dulbecco s modified eagle medium, by Lonza (5 mentions) Dmem f12, by Thermo Fisher Scientific (5 mentions) Penicillin streptomycin mixture, by Lonza (4 mentions) Rpmi 1640 medium, by Lonza (4 mentions) Collagenase 4, by Merck Group (4 mentions) Egm 2, by Lonza (4 mentions)

163 protocols using trypsin

Culturing Melanoma Cell Lines WM115 and G361

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WM115 cells originating from a primary melanoma (received as a generous gift from Piia Takabe, MSc, and Docent Sanna Pasonen-Seppänen, PhD, Institute of Biomedicine, University of Eastern Finland) were cultured in High Glucose DMEM (PAA Cell Culture Company, Cambridge, UK) supplemented with 5 % inactivated fetal bovine serum (FBS) (PAA Cell Culture Company), 2 mM glutamine (Lonza, Basel, Switzerland), 1 mM sodium pyruvate (Thermo Fisher Scientific, Waltham, MA, USA) and 50 U/mL penicillin – 50 µg/mL streptomycin sulfate (Lonza). Cells were passaged twice a week at a 1:10 split ratio using 0.02% trypsin (w/v) - 0.02% EDTA (w/v) (Lonza).
G361 cells originating from a primary melanoma (Sigma-Aldrich, Munich, Germany) were cultured in McCoy’s 5a medium (Sigma-Aldrich, order No. 88030401) supplemented with 2 mM glutamine (Lonza) and 10% FBS (PAA Cell Culture Company). Cells were passaged twice a week at a 1:10 split ratio using 0.02% trypsin (w/v) - 0.02% EDTA (w/v) (Lonza).

Siiskonen H, & Harvima I.T. (2022). Mast cell chymase is in contact with melanoma cells in vivo and it detaches melanoma cells from the substratum in vitro. Translational Cancer Research, 11(12), 4315-4325.

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Expansion of Human Mesenchymal Stem Cells

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hMSCs were purchased from Lonza (Basel, Switzerland) and cultured in MF-medium (TOYOBO, Tokyo, Japan). For the maintenance of hMSCs, the culture media were replenished every 2 days. hMSCs were detached and dissociated into single cell suspension by 4–5 min incubation with Trypsin solution [Trypsin/ethylenediaminetetraacetic acid (EDTA) for Mesenchymal Stem Cells, Lonza]. Live cell numbers were manually counted using hemocytometer.

Osada H., Kawatou M., Takeda M., Jo J.I., Murakami T., Tabata Y., Minatoya K., Yamashita J.K, & Masumoto H. (2020). Accuracy of spiked cell counting methods for designing a pre-clinical tumorigenicity study model. Heliyon, 6(7), e04423.

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Culturing Mucoepidermoid Carcinoma Spheroids

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Non-adherent spheroids derived from mucoepidermoid carcinoma cells (salispheres) were cultured in ultralow attachment 6-well plates (Corning; Corning, NY, USA) for 10 days with DMEM-F12 (Invitrogen, Carlsbad, CA, USA) supplemented with 1% N-2 supplement (Invitrogen), 1% GlutaMAX (Invitrogen), 1% penicillin/streptomycin (Invitrogen), 1 μM dexamethasone (MilliporeSigma), 10 μg/ml insulin (MilliporeSigma), 20 ng/ml EGF (MilliporeSigma), and 20 ng/ml basic fibroblast growth factor (bFGF; MilliporeSigma). For in vitro passaging, salispheres were exposed to 0.25% trypsin (MilliporeSigma) for 5 min, and then mechanically dissociated. A neutralizing solution (TNS; Lonza) was used to neutralize trypsin. Cells were counted and then added to new 6-well ultralow attachment plates (Corning; Corning, NY, USA). Colonies of 50 cells or more were considered salispheres.

Andrade N.P., Warner K.A., Zhang Z., Pearson A.T., Mantesso A., Guimaraēs D.M., Altemani A., Mariano F.V., Nunes F.D, & Nör J.E. (2021). Survival of salivary gland cancer stem cells requires mTOR signaling. Cell Death & Disease, 12(1), 108.

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Passaging A549 Cells with Trypsin

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A549 cells were passaged when they were 80% confluent, about every third day. Media was removed by aspiration and 5 ml of phosphate buffered saline pH 7.2 (Lonza Verviers SPRL, Belgium) were added to wash adherent cells. To detach the adherent cells, 1 ml of 2.5% (w/v) Trypsin (Lonza Verviers SPRL, Belgium) was added to the T-25 flasks and cells were incubated for 5 min at 37 °C. Then, cells were detached and observed under the inverted microscope (Micro master inverted digital microscope, Thermo Fisher Scientific Inc., USA) every 2–3 min.

El-Kishky A.H., Moussa N., Helmy M.W, & Haroun M. (2022). GANT61/BI-847325 combination: a new hope in lung cancer treatment. Medical Oncology (Northwood, London, England), 39(10), 144.

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Cell Culture of Common Cell Lines

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All cell lines were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA)
and resulted negative for mycoplasma test (MycoAlert TM Mycoplasma Detection Kit, Lonza Sales AG, Verviers, Belgium). Murine fibroblasts (NIH/3T3), melanoma (B16.F10) and human umbilical vein endothelial (HUVEC) cells were cultured as monolayers at 37°C in a 5% CO 2 -containing humidified atmosphere in Dulbecco's modified Eagle's medium (DMEM). The medium was supplemented with 10% (v/v) of heat-inactivated foetal bovine serum, 2 mM of Glutamine, 100 U/ml of penicillin and 100 μg/ml of streptomycin. At 80% confluency, cells were trypsinized with 0.01% trypsin and 0.02% EDTA in Phosphate-buffered Saline (PBS). All cells employed for cellular tests were at passages included between 5 and 10. Culture medium, fetal bovine serum, penicillin-streptomycin mixture and trypsin were all purchased from Lonza (Lonza Sales AG, Verviers, Belgium).

Filippi M., Catanzaro V., Patrucco D., Botta M., Tei L, & Terreno E. (2017). First in vivo MRI study on theranostic dendrimersomes. Journal of controlled release : official journal of the Controlled Release Society, 248.

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Culturing Mouse Melanoma B16-F10 Cells

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B16‐F10 (mouse melanoma cells) were obtained from American Type Culture Collection. B16‐F10 cells were cultured in EMEM supplemented with 10% FBS, 100 μg/mL penicillin and 100 μg/mL streptomycin. The cells were grown at 37°C in a humidified atmosphere containing 5% CO₂. At confluence, cells were detached by adding 1 mL of Trypsin–EDTA solution (0.25%_w/v Trypsin, 0.53 mM EDTA). EMEM, FBS and Trypsin were purchased from Lonza (Verviers, Belgium). The penicillin–streptomycin mixture was purchased from Sigma Chemical Co. (St. Louis, MO, USA).

Anemone A., Capozza M., Arena F., Zullino S., Bardini P., Terreno E., Longo D.L, & Aime S. (2021). In vitro and in vivo comparison of MRI chemical exchange saturation transfer (CEST) properties between native glucose and 3‐O‐Methyl‐D‐glucose in a murine tumor model. Nmr in Biomedicine, 34(12), e4602.

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HepG2 Cell Culture and Hydrogel Embedding

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Two
vials containing approximately 2 million HepG2 cells at passage 99
were kindly provided by “Istituto Zooprofilattico Sperimentale
della Lombardia e dell’ Emilia-Romagna”. Cells were
thawed, counted in a hemocytometer by means of the trypan blue (Sigma-Aldrich,
Lot. no. RNBD9396, US) exclusion assay, and seeded in a 6-well plate
with a seeding ratio of approximately 300,000 cells/well. The medium
used to culture cells was composed of EMEM with Earl’s salts
(EuroClone, Cat. no. ECB2071L, IT), 10% (v/v) FBS (EuroClone, Lot.
no. EU-S021179, IT), 1% (v/v) Na pyruvate (EuroClone, Lot. no. EU-M00QU,
IT), 1% (v/v) glutamine (EuroClone, Lot. no. EU-M0150017, IT) and
1% (v/v) penicillin–streptomycin (Lonza, Lot. no. 2MB027, BE),
and was refreshed every 48 h. Cells were expanded up to passage 6
and then cryopreserved (10⁶ cells/cryovial) in complete
medium + 10% (v/v) DMSO (Sigma-Aldrich, Lot. no. SZBD2870V, US). To
be embedded within hydrogels, cells were expanded, detached with trypsin
(Lonza, Lot. no. 4MB146, BE), counted, suspended in a fresh medium,
and embedded within gels as described in 2.3 at a final density of
3 × 10⁶ cells/gel. Hydrogels were then submerged with
2 mL of fresh medium and incubated at 37 °C, 95% humidity, and
5% CO_2.

Guagliano G., Volpini C., Sardelli L., Bloise N., Briatico-Vangosa F., Cornaglia A.I., Dotti S., Villa R., Visai L, & Petrini P. (2022). Hep3Gel: A Shape-Shifting Extracellular Matrix-Based, Three-Dimensional Liver Model Adaptable to Different Culture Systems. ACS Biomaterials Science & Engineering, 9(1), 211-229.

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Cell Culture of Jurkat and HEK293G Lines

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Jurkat cells (clone E6-1) were maintained in T175 flasks containing RPMI 1640 medium (Lonza) supplemented with 10% fetal calf serum (FCS; Sigma Aldrich) and 2 mM L-glutamine (Sigma Aldrich) at 37°C/5% CO₂. Fresh medium was added to the cells every 2-3 days. Cells were passaged at 70% confluency by withdrawing 2.5 ml of the cells into a fresh T175 flask with medium. HEK293G cells (Glosensor cAMP HEK293, Promega) were grown in T75 flasks containing 25 ml Dulbecco’s Modified Eagle’s Medium (DMEM; Sigma Aldrich, UK) supplemented with 10% FCS at 37°C/5% CO₂. Cells were passaged at 70-80% confluency using phosphate buffered saline (PBS; Lonza) and trypsin (0.25% w/v in versene; Lonza). A mixed population HEK293G cell line was created by transfecting cells with the LgBiT-CXCR4 construct using FuGENE® (Promega) according to the manufacturer’s instructions, followed by selective pressure (1 mg/ml G418) for two to three weeks. HEK293G cells stably expressing the SNAP-CXCR4 construct were kindly gifted from Dr. J. Goulding (University of Nottingham).

Soave M., Heukers R., Kellam B., Woolard J., Smit M.J., Briddon S.J, & Hill S.J. (2020). Monitoring Allosteric Interactions with CXCR4 Using NanoBiT Conjugated Nanobodies. Cell Chemical Biology, 27(10), 1250-1261.e5.

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STRO-1+ Stem Cell Culture and Osteogenic Differentiation

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STRO-1+–enriched SSCs were maintained at 5% CO₂ at 37°C in culture
flasks and used at passage 1. Cell culture media used was basal culture media
unless otherwise stated. Osteogenic media was basal media supplemented with
10 nM vitamin D₃ (Sigma) and 100 μM ascorbic acid (Sigma). Prior to
seeding, cells were treated with collagenase IV (Sigma) and released from the
flask using 1× trypsin (Lonza). STRO-1+ SSC were seeded into 24-well plates at
1000 cells/cm². After 24 h of seeding, surfaces were transferred
to new 24-well plate and media replaced. Media was subsequently changed every
3–4 days for the duration of the experiments.

Waddell S.J., de Andrés M.C., Tsimbouri P.M., Alakpa E.V., Cusack M., Dalby M.J, & Oreffo R.O. (2018). Biomimetic oyster shell–replicated topography alters the behaviour of human skeletal stem cells. Journal of Tissue Engineering, 9, 2041731418794007.

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Quantifying 3D Cancer Cell Morphogenesis

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Morphogenesis assays were performed by mixing cells with ECM (BD Biosciences Matrigel catalog #356231 or Trevigen Cultrex catalog #3443-005-01) on ice, plated in triplicate into 24-well tissue culture plates, and incubated at 37 °C and 5% CO₂. After solidification for 30 min, culture media was added. For coculture experiments, CAFs were either overlaid on top of solidified Cultrex matrix or coembedded with TUM622 at the time of plating at a ratio of 2:1. Cultures were kept for 8 d to 12 d, with media replaced every 2 d. Serial passaging of TUM622 acini was performed by extracting acini (day 10) from the ECM via Cultrex 3D-cell harvesting kit (catalog #3448-020-K; Trevigen) according to manufacturer’s recommendation. Acini were made into a single-cell suspension with trypsin (catalog #CC-5034; Lonza) and replated as described above. Inhibition of Notch and Wnt signaling pathway in 3D cultures was performed with small-molecule inhibitors/agonist IWR-1 (catalog #681669; EMD Millipore), CHIR-99021 (catalog #SML1046; Sigma) and DBZ (catalog #4489; Tocris) at the indicated concentrations. Media containing inhibitors/agonists were replaced once every 2 d for 10 d. The number and size of acini/spheroids were quantified with GelCount (Oxford Optronix, Inc.).

Chen S., Giannakou A., Wyman S., Gruzas J., Golas J., Zhong W., Loreth C., Sridharan L., Yamin T.T., Damelin M, & Geles K.G. (2018). Cancer-associated fibroblasts suppress SOX2-induced dysplasia in a lung squamous cancer coculture. Proceedings of the National Academy of Sciences of the United States of America, 115(50), E11671-E11680.

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