Melanoma cells (SK‐Mel‐28, SK‐Mel‐147, 451LU, WM902B, B16R2L, and B16‐F10), obtained from (ATCC), were cultured in DMEM (Invitrogen) supplemented with 10% FBS. All cell lines were authenticated using The GenePrint 10 System (Promega, MA). Cells were tested for mycoplasma contamination (Mycoplasma Detection Kit (LT07‐318), Lonza, Basel, Switzerland) regularly and before injection in mice. When indicated, cells were stably infected with mCherry pLV‐puro lentiviral vectors as described before (Tormo et al, 2009 (link)).
Geneprint 10 system
Manufactured by Promega
Sourced in United States, Spain, Italy, Australia, United Kingdom, Taiwan, Province of China, Canada
The GenePrint 10 System is a multiplex short tandem repeat (STR) amplification kit designed for human identification applications. It simultaneously amplifies ten autosomal STR loci and the Amelogenin gender-determining marker in a single PCR reaction. The kit provides a streamlined workflow for generating high-quality, robust STR profiles from a variety of sample types.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (47 mentions)
Mycoalert mycoplasma detection kit, by Lonza (24 mentions)
Streptomycin, by Thermo Fisher Scientific (24 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (23 mentions)
Penicillin, by Thermo Fisher Scientific (22 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (15 mentions)
Rpmi 1640, by Thermo Fisher Scientific (14 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (13 mentions)
Fetal bovine serum (fbs), by Merck Group (8 mentions)
Glutamine, by Thermo Fisher Scientific (8 mentions)
L glutamine, by Thermo Fisher Scientific (7 mentions)
Mcf 7, by American Type Culture Collection (7 mentions)
Mycoalert, by Lonza (7 mentions)
Hydrocortisone, by Merck Group (7 mentions)
Rpmi 1640, by Lonza (7 mentions)
Mda mb 231, by American Type Culture Collection (7 mentions)
Ezkill mycoplasma removal reagent, by HiMedia (6 mentions)
Genemarker hid, by SoftGenetics (6 mentions)
Dmem f12, by Thermo Fisher Scientific (6 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (6 mentions)
310 protocols using geneprint 10 system
1
Melanoma Cell Culture and Modification
2
Culturing Human Breast Cancer Cell Lines
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The MDA-MB-468 human breast cancer cell line was obtained from The Brisbane Breast Bank, UQCCR, Brisbane, Australia, and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) with high glucose (D6546; Sigma-Aldrich, St Louis, MO, USA), supplemented with 10% fetal bovine serum (FBS) and 4 mM L-glutamine (25030; Thermo Fisher Scientific, Waltham, MA, USA). PMC42LA human breast cancer cell line was obtained from Dr Leigh Auckland, Deakin University, Melbourne, Australia [31 (link),32 (link)], and maintained in Roswell Park Memorial Institute (RPMI)-1640 medium (11875085; Life Technologies, Carlsbad, CA, USA) with 10% FBS. Cells were maintained at 37 °C and 5% CO2 in a humidified incubator. For hypoxia experiments, cells were placed in a hypoxic incubator (37 °C, 1% O2, 5% CO2, and 94% N2) for durations stated in the results. Cells were examined for mycoplasma every six months using MycoAlert kit (Lonza, Basel, Switzerland) and validated by short tandem repeat (STR) profiling using The GenePrint® 10 System (Promega, Madison, WA, USA) at the Queensland Institute of Medical Research (QIMR) Berghofer Medical Research Institute, Brisbane, Australia.
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Breast Cancer Cell Line Characterization
4
Prostate Cancer Cell Line Validation
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PC-3 (CRL-1435) human prostate cancer cell line was purchased from ATCC (Manassas, VA) and maintained in the recommended culture media. The identity of the cell line was checked by Cell ID System and Promega GenePrint 10 System through short tandem repeat analysis (Mission Biotech, Taipei, Taiwan). A comprehensive methods section is available in Supplementary Methods .
5
Cell Lines Maintenance and Characterization
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Ovarian cancer cell lines OVCAR4 (NCI, Frederick, MA), TOV21G (Fran Balkwill, Barts Cancer Institute, London, UK), erythroleukemia cell line K562 (Vignir Helgason, University of Glasgow, Glasgow, UK), and human NK cell line NK-92 (ATCC, Manassas, VA) were incubated at 37°C in 5% CO2. OVCAR4 and TOV21G were maintained in DMEM with 10% FBS, 2 mM L-Glutamine, and 100 μg/mL penicillin/streptomycin. NK-92 cells were maintained in MEM-alpha with 12.5% FBS, 12.5% horse serum, 2 mM L-Glutamine, and 5 ng/mL interleukin-2 (IL-2). K562 were maintained in RPMI with 10% FBS plus 2 mM L-Glutamine, and 100 μg/mL penicillin/streptomycin. All lines were tested regularly for mycoplasma infection. All human cancer cell lines were verified by short tandem repeat profiling at the Cancer Research UK Beatson Institute using the Promega GenePrint 10 system (Promega, Southampton, UK). Human NK cells were isolated, resuspended in RPMI with 10% FBS plus 2 mM L-Glutamine and 100 μg/mL penicillin/streptomycin, and used immediately without additional IL-2 or IL-15.
6
Prostate Cancer Cell Line Validation
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PC‐3 (CRL‐1435) and DU145 (HTB‐81) human prostate cancer cell lines were purchased from ATCC (Manassas, VA) and maintained in the recommended culture media. The identity of the cell line was checked by Cell ID System and Promega GenePrint 10 System through short tandem repeat analysis (Mission Biotech, Taipei, Taiwan), and passage numbers used for the experiments are <25. A comprehensive methods section is available in Supplementary methods.
7
Cell Line Characterization and Preparation
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Cell lines ES2 and TOV21G were obtained from the American Type Tissue Collection (ATCC). HCT116 isogenic ARID1A (Q456*/Q456*) and parental lines were purchased from Horizon Discovery (Cambridge, UK). These were developed by knock‐in of a premature stop codon (Q456*). Cell lines were cultured in a humidified 37 °C incubator with 5% CO2. Cell lines were tested to confirm no mycoplasma infection using Mycoalert™ Mycoplasma Detection Kit as per manufacturer's instructions (Lonza, Slough, UK). Cell line identity was confirmed with short tandem repeat typing using the Promega GenePrint®10 system (Promega, Southampton, UK). Cell pellets were formalin fixed and paraffin‐wax embedded (FFPE) for antibody optimisation.
8
Establishing 5-FU-Resistant Colorectal Cancer Cell Lines
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The CRC patient samples were obtained at the First Affiliated Hospital of Sun Yat-sen University and were approved by the institutional review board of the hospital. The study was compliant with all relevant ethical regulations regarding research involving human participants. HCT-8, HCT-116, HEK-293T, and MC38 cells were obtained from the Procell. All cells were cultured in RPMI-1640 or DMEM (Corning, USA) with 10% fetal bovine serum at 37°C in 5% CO2. All cell lines were stored in multiple back up upon receipt to reduce risk of phonotypic drift, and tested with mycoplasma free by Mycoplasma Stain Assay Kit (C0296, Beyotime, China). Cell line authentication was validated by the STR analysis using GenePrint 10 System according to the manufacturer’s instruction (B9510, Promega, USA). 5-FU-resistant HCT-8 and HCT116 cell lines were generated by exposing the cells with 5-FU at 5% of IC concentration for 3 days and gradually increased the concentration by 5% of IC until reaching the IC50.
9
Characterization of GBM Cell Lines
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All reagents were purchased from Sigma-Aldrich unless otherwise stated. Doxycline (Dox) was diluted to a concentration of 1μg/ml in cell culture medium as a working concentration. The human glioblastoma (GBM) cell lines U87 were originally purchased from ATCC (Manassas, VA). GBM neurosphere culture (HSR-GBM1A) were originally established by Vescovi and colleagues15 (link) and further characterized by us.16 (link)–18 (link) Both cells lines are free from mycoplasma and authenticated with short tandem repeat (STR) profiling by Johns Hopkins Genetic Resources Core facility using Promega GenePrint 10 system (Madison, WI). U87 cells were cultured in Minimum Essential Media (MEM, Thermo Fisher Scientific, Grand Island, NY) supplemented with sodium pyruvate (1%), sodium bicarbonate (2%), non-essential amino acid (1%) and 10% fetal calf serum (FCS, Gemini Bio-products, West Sacramento, CA). HSR-GBM1A (GBM1A) cells contain CD133+ GBM stem-like cells and form infiltrative orthotropic xenografts that have been extensively characterized by others and our group.19 (link),20 (link) GBM1A neurospheres were cultured in DMEM/F12 medium supplemented with epidermal growth factor (EGF) (Peprotech, Rocky Hill, NJ) and fibroblast growth factor (FGF) (Peprotech). Cells were incubated in a humidified incubator containing 5% CO2/95% air at 37°C, and passaged every 4-5 days.
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Characterization of GBM Cell Lines
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