The human lung cancer A549, colon cancer SW620, osteosarcoma cancer HOS and 143B cell lines were purchased from the American Type Culture Collection (ATCC). The human prostate cancer PC-3U cells represents a clone from the original PC-3 cell line (ATCC CRL-1435)54 (link). The human glioblastoma U251 MG (formerly U-373 MG) originated from the European Collection of Authenticated Cell Cultures (ECACC). The four cancer cell lines stably expressing GFP; U251-GFP, SW620-GFP, PC-3U-GFP, and A549-GFP, were cultured in RPMI media supplemented with 10% (v/v) FBS, and 1% (v/v) PS (Gibco). Cells were cultured at 37 °C in the presence of 5% (v/v) CO2. Cancer cells were maintained under passage 15. Mycoplasma test (Eurofins GATC Biotech) confirmed that the cancer cells were free from mycoplasma infection, and the identity of the cancer cell lines was verified by STR profiling (Microsynth).
U 373 mg
Manufactured by European Collection of Authenticated Cell Cultures
Sourced in United States, United Kingdom, Sweden
U-373 MG is a glioblastoma cell line derived from a malignant human brain tumor. It is a well-characterized cell line commonly used in brain cancer research.
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Lab products found in correlation
U 251 mg, by European Collection of Authenticated Cell Cultures (4 mentions)
Sw620, by Thermo Fisher Scientific (1 mentions)
Sw620, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ln229, by American Type Culture Collection (1 mentions)
Powerplex hs genotyping kit, by Promega (1 mentions)
U 87 mg, by European Collection of Authenticated Cell Cultures (1 mentions)
3500 genetic analyzer, by Thermo Fisher Scientific (1 mentions)
5 protocols using u 373 mg
1
Characterization of Cancer Cell Lines
2
Culturing Human Glioblastoma, Astrocytes, and Tracheal Epithelial Cells
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Human glioblastoma cell line U-251MG (cat no. 09063001, formerly U-373 MG) was obtained from the European Collection of Authenticated Cell Cultures and cultured in Minimum Essential Medium Eagle (EMEM, Gibco, Grand Island, NY, USA) + 2 mM glutamine + 1% nonessential amino acids + 1 mM sodium pyruvate (NaP) + 10% fetal bovine serum (FBS; Gibco) at 37°C in a 5% CO2 humidified atmosphere. Human astrocytes (HAs) were obtained from iXCells (San Diego, CA, USA) and cultured in an astrocyte medium (cat no. 10HU-035, iXCells) at 37°C in a 5% CO2 humidified atmosphere. Human tracheal epithelial cells (TECs) were obtained from ATTC (PCS-300-013, USA) and cultured in an airway epithelial cell basal medium (cat no. PCS-300-030, Sigma) at 37°C in a 5% CO2 humidified atmosphere.
3
Culturing U-251 Glioblastoma and Vero Cells
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Human U-251 glioblastoma astrocytoma [U-251 MG (formerly known as U-373 MG; European Collection of Authenticated Cell Cultures – ECACC 09063001)] were cultured in Dulbecco’s modified Eagle’s/Nutrient Mixture F-12 (DMEM/F-12) medium. Monkey Vero cells (ATCC® # CCL-81TM) were cultured in Dulbecco’s modified Eagle’s medium (DMEM). Both media were supplemented with 10% Fetal Bovine Serum (FBS), non-essential amino acids, 2 mM L -glutamine, and sodium pyruvate, respectively, and cells were maintained at 37°C in 5% CO2. Cells were trypsinized and passaged every 2 – 3 days.
4
Characterization of Human Glioblastoma Cell Lines
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The human glioblastoma cell lines LN229 (ATCC-CRL-2611), U251 (ECACC 89081403; formerly known as U373MG), U87MG (ECACC 89081402), and T98G (ECACC No. 92090213) were obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK) or the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured as described previously [24 (link)]. Human primary GBM cultures (n = 2) were produced by dissociation and cultured according to established techniques as described before [24 (link)]. Human primary GBM stem-like cell cultures (n = 8) as well as GBM cell line-derived stem-like cells were established and intensively characterized by the formation of neurospheres, the ability to survive and proliferate under stem cell conditions, and the ability to differentiate into more mature cells as described before [3 (link),25 (link),26 (link),27 (link)]. The purity of the GBM cells was ascertained by immunostaining with cell type-specific markers and by the absence of contamination with mycoplasms. GBM cell line identity was verified by short tandem repeat profiling at the Department of Forensic Medicine (Kiel, Germany) using the Powerplex HS Genotyping Kit (Promega, Madison, WI, USA) and the 3500 Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA, USA) as previously described [3 (link)].
5
Culturing Human Glioblastoma Cells
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Human glioblastoma astrocytoma (U251) cells [U251 MG (previously known as U-373 MG; European Collection of Authenticated Cell Cultures—ECACC 09063001, Sweden)] were grown in Dulbecco’s modified Eagle’s/Nutrient Mixture F-12 (DMEM/F-12) media completed with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1× non-essential amino acids and 1× sodium pyruvate at 37 °C in 5% CO2. Cells were trypsinized every 2–3 days for passage. Viral titers were determined using plaque assays in Vero cells, as described [54 (link)].
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