Trypsin edta 1

Manufactured by Lonza

Sourced in Netherlands, Belgium

Trypsin EDTA 1x is a cell culture reagent used for the dissociation and detachment of adherent cells from cell culture surfaces. It is a mixture of the proteolytic enzyme trypsin and the chelating agent EDTA, which together facilitate the breakdown of cell-cell and cell-substrate adhesions, allowing cells to be harvested and subcultured.

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Lab products found in correlation

Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Lonza (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Foetal bovine serum (fbs), by Bovogen (1 mentions) C39211, by PromoCell (1 mentions)

Spelling variants of this product

Trypsin-EDTA (248 citations) Trypsin (163 citations)

3 protocols using trypsin edta 1

Culturing MDA-MB-231 Breast Cancer Cells

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MDA-MB-231 breast cancer cells (A kind gift from Dr. Dana Mustafa, Erasmus Medical Center, Rotterdam, the Netherlands) were cultured in the culture medium which was RPMI 1640 medium (Gibco, Thermofisher Scientific, the Netherlands) supplemented with 10% (v/v) foetal bovine serum (FBS, Bovogen Biologicals, Australia) and 1% (v/v) penicillin-Streptomycin (Lonza, Westburg, the Netherlands) at 37^°C and 5% CO₂. After more than 80% confluency, the cells were removed from the bottom of the culture flask using Trypsin EDTA 1× (Lonza, Westburg, the Netherlands) and centrifuged in the growth medium at 900 rpm for 5 min. The supernatant was then removed and new culture medium was added to the tube.
For chemotaxis experiments, recombinant human epidermal growth factor (EGF, Gibco, Thermo Fisher Scientific) was diluted to 50 ng/ml in the culture medium (chemotactic medium). Before chemotaxis, cells were starved in the starving medium (RMPI 1640, 1% (v/v) penicillin-Streptomycin and 0.1% bovine serum albumin) for 2 h.

Eslami Amirabadi H., SahebAli S., Frimat J.P., Luttge R, & den Toonder J.M. (2017). A novel method to understand tumor cell invasion: integrating extracellular matrix mimicking layers in microfluidic chips by “selective curing”. Biomedical Microdevices, 19(4), 92.

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Isolation of Primary Human Umbilical Vein Endothelial Cells

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Primary HUVECs were isolated from human umbilical cords obtained at the Leiden University Medical Center after written informed consent and ensuring that collection and processing of the umbilical cord was performed anonymously. The umbilical vein was flushed with PBS, using glass cannulas, to remove all remaining blood. Endothelial cells were detached by infusion of the vein with Trypsin/EDTA (1×) (BE02‐007E, Lonza, Verviers, Belgium) solution and incubation at 37°C for 15 min. After incubation, the cell suspension was collected and taken up in endothelial cell growth medium (EGM2 medium, C222111 supplemented with C39211, Promocell, Heidelberg, Germany) with 1% antibiotics. After flushing the umbilical vein once more with PBS, to ensure all detached cells are collected, cells were pelleted by centrifugation at 1200 rpm for 7 min. The cell pellet was dissolved in fresh EGM2 medium and cells were cultured on gelatin (1%) coated surfaces.

Vreeken D., Bruikman C.S., Cox S.M., Zhang H., Lalai R., Koudijs A., van Zonneveld A.J., Hovingh G.K, & van Gils J.M. (2020). EPH receptor B2 stimulates human monocyte adhesion and migration independently of its EphrinB ligands. Journal of Leukocyte Biology, 108(3), 999-1011.

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Isolation of Primary Human Umbilical Vein Endothelial Cells

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Primary human umbilical vein endothelial cells (HUVECs) were isolated from human umbilical cords obtained at the Leiden University Medical Center after written informed consent and ensuring that collection and processing of the umbilical cord was performed anonymously. The umbilical vein was flushed with PBS, using glass cannulas, to remove all remaining blood. Endothelial cells were detached by infusion of the vein with Trypsin/EDTA (1×, BE02-007E; Lonza) solution and incubation at 37ºC for 15 minutes. After incubation, the cell suspension was collected and taken up in endothelial cell growth medium (Promocell; C222111 supplemented with C39211) with 1% antibiotics. After flushing the umbilical vein one more with PBS, to ensure all detached cells are collected, cells were pelleted by centrifugation at 1200 rpm for 7 minutes. Cell pellet was dissolved and maintained in EGM2 medium, and cells were cultured on gelatin (1%) coated surfaces.

Bruikman C.S., Vreeken D., Hoogeveen R.M., Bom M.J., Danad I., Pinto-Sietsma S.J., van Zonneveld A.J., Knaapen P., Hovingh G.K., Stroes E.S.G, & van Gils J.M. (2020). Netrin-1 and the Grade of Atherosclerosis Are Inversely Correlated in Humans. Arteriosclerosis, thrombosis, and vascular biology, 40(2).

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