HEK293FT (RRID:CVCL_6911 ), MDA-MB-231 (RRID:CVCL_0062 ), MDA-MB-468 (RRID:CVCL_0419 ), MDA-MB-436 (RRID:CVCL_0623 ) and COV362 (RRID:CVCL_2420 ), cells were cultured in high-glucose Dulbecco's Modified Eagle Medium (DMEM) containing HEPES buffer, penicillin, and streptomycin (Wisent cat: 319-005-CL) supplemented with 10% fetal bovine serum (FBS) (Wisent cat: 080-150). OVCAR8 (RRID:CVCL_1629 ) cells were cultured in high-glucose Roswell Park Memorial Institute's (RPMI) medium containing HEPES buffer, penicillin, and streptomycin (Wisent cat: 350-700-CL) supplemented with 10% FBS (Wisent cat: 080-150). SUM149PT (RRID:CVCL_3422 ), cells were cultured in Ham's F-12 (F12) medium mixture (Gibco cat: 31765035) supplemented with 5% FBS, 10 mM HEPES (Gibco cat: 15630080) 1 μg/ml hydrocortisone (Sigma cat: H0888) and 5 μg/ml insulin (Sigma cat: I-1882). MCF10A (RRID:CVCL_0598 ), cells were grown in a DMEM/F12 medium mixture (Gibco cat: 11320033) supplemented with 5% horse serum (Invitrogen: 16050-122), 20 ng/ml epidermal growth factor (EGF), 100 ng/ml Cholera toxin, 0.5 mg/ml hydrocortisone (Sigma cat: H0888) and 10 μg/ml insulin (Sigma cat: I-1882). Cell lines were passaged using 0.05% Trypsin-EDTA solution (Wisent cat: 325-542-EL) and grown in a humidified atmosphere at 37°C and 5% CO2.
Trypsin edta solution
Manufactured by Wisent
Sourced in Canada
Trypsin/EDTA solution is a cell culture reagent used to dissociate and detach adherent cells from a culture surface. It contains the enzyme trypsin, which cleaves the peptide bonds in the proteins that hold cells together, and EDTA, which chelates calcium and magnesium ions to facilitate cell detachment.
Automatically generated - may contain errors
Lab products found in correlation
Mda mb 231, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Wisent (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Mtt powder, by Merck Group (2 mentions)
Hydrocortisone, by Thermo Fisher Scientific (1 mentions)
Insulin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Wisent (1 mentions)
Streptomycin, by Wisent (1 mentions)
Calcium chloride cacl2, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Penicillin streptomycin antibiotic, by Thermo Fisher Scientific (1 mentions)
Alginic acid sodium salt, by Merck Group (1 mentions)
Live dead cell viability assay kit, by Merck Group (1 mentions)
Hoechst 33342 nuclei dye, by Merck Group (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
4 protocols using trypsin edta solution
1
Cell Culture Protocols for Cancer Cell Lines
2
Cytotoxicity Evaluation of MDA-MB-231 Cells
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The alginic acid Sodium salt, dimethyl sulfoxide (DMSO), Sodium chloride, Calcium chloride (CaCl2) and gelatin from bovine skin (type B) were obtained from Sigma-Aldrich (Canada). For cell culture studies, MDA-MB-231 were purchased from ATCC, DMEM (Dulbecco's Modified Eagle Medium), FBS (fetal bovine serum), penicillin/streptomycin, Trypsin/EDTA solution at 0.25% (w./v.) and phosphate buffer saline (PBS) tablets were bought from Wisent Bioproducts. (3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide) MTT powder, Triton X-100, BSA (bovine serum albumin) and paraformaldehyde were bought from Sigma-Aldrich (Canada). Furthermore, a Live/Dead Cell Viability assay kit (CBA415), Hoechst 33342Nuclei Dye were provided from Sigma-Aldrich (Canada). Additionally, Anti-Ki67 antibody (ab15580), Alexa Fluor 546 Goat anti-Rabbit IgG (H + L) were purchased from Abcam and Invitrogen (Canada), respectively.
3
Establishing MDA-MB-231 Breast Cancer Xenografts
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In this study, MDA-MB-231 breast cancer cell lines were used to grow tumours in the hind leg of female severe combined immunodeficiency (SCID)-B17 mice obtained from Charles River Canada. Cells obtained from the American Type Culture Collections (ATCC, MD, USA) were incubated at 37 °C in 5% CO2 in RPMI-1640 medium supplemented with 10% fetal bovine serum (Sigma-Aldrich) and 1% penicillin/streptomycin antibiotics (ThermoFisher Scientific). On reaching 80% confluency, a 0.05% Trypsin–EDTA solution (Wisent BioProducts) was used to trypsinize the cells in preparation for injection. For tumour induction, 5 × 106 cells/100 µL cells were re-suspended in Mg+/Ca+ Dulbecco's phosphate-buffered saline and were injected in the hind limb of the mice. It took about 4–6 weeks of xenografts to grow 5–10 mm and ready for treatment. A mixture of ketamine (100 mg/kg) andxylazine (5 mg/kg) was used to anesthetize animals during the injection of the cells and the therapy procedure. Throughout the experiments, mice were closely monitored by placing the heat lamps and heating pads nearer to prevent irregular body temperature. Each treatment group consisted of 4 or more than 4 animals.
4
Bioprinting of Breast Cancer Cell Lines
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Gelatin from bovine skin (type B) and alginic acid sodium salt, dimethyl sulfoxide (DMSO), calcium chloride, and sodium chloride powder were purchased from Sigma-Aldrich (Canada). calcium chloride was dissolved in Millipore water at a final concentration of 4%, filtered with 0.2 u filters and stored in a sterile condition for further use. For cell culture studies, MCF7 and MDA-MB-231 cell lines were purchased from ATCC (Cedarlane, Canada). DMEM (Dulbecco’s Modified Eagle’s Medium, without sodium pyruvate, with 4.5 g/l glucose, with L-glutamine), PBS (phosphate buffer saline) 1×, Penicillin-streptomycin, Trypsin/EDTA solution at 0.25% (w./v.) and FBS (fetal bovine serum) were purchased from Wisent Bioproducts (Canada). (3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide) MTT powder, Triton X-100, BSA (bovine serum albumin), paraformaldehyde, a Live/Dead Cell Viability assay kit (CBA415) and Hoechst 33342 Nuclei Dye, Propidium Iodide (PI) and Calcine were bought from Sigma-Aldrich (Canada). For bioprinting, the mixing syringe (5ml) with the mixing ratio of 4:1, and compatible mixer tips were purchased from Sulzer Inc. (Switzerland). Empty cartridges and nozzles purchased from Cellink (Sweden)
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