Umbilical Cord Blood (CB) and Adult Blood (AB) samples were collected through the Basque Biobank (http://www.biobancovasco.org ) under an institutional review board-approved protocol by the Basque Committee of Ethics and Clinical Research. The methods were carried out in accordance with the approved guidelines. The Basque Biobank complies with the quality management, traceability and biosecurity, set out in the Spanish Law 14/2007 of Biomedical Research and in the Royal Decree 1716/2011. All study subjects were provided written informed consent. CB units that contain between 1.5 × 109 and 8 × 108 mononuclear cells were used for research purposes24 (link). K562 was purchased from ATCC (CCL-243). Nalm-6 cell line was provided by the Immunotherapy Department of the Hospital Clinic-IDIBAPS, Barcelona. Acute Lymphoblastic Leukemia (ALL) cells (GM20390 and GM16726) were purchased from Coriell Company. All cell lines were cultured with RPMI, 10% FBS, 1% penicillin/streptomycin, 1% Glutamax, 1% NEAA, and 1% sodium pyruvate.
Ccl 243
Manufactured by American Type Culture Collection
Sourced in United States
The CCL-243 is a cell line derived from human prostate cancer tissue. It is a widely used tool in cancer research for studying prostate cancer biology and evaluating potential therapeutic agents.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Ccl 185, by American Type Culture Collection (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Crl 1593, by American Type Culture Collection (2 mentions)
Ccl 119, by American Type Culture Collection (2 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
Tgf β, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Rpmi medium, by Thermo Fisher Scientific (1 mentions)
Hemin, by Merck Group (1 mentions)
Amaxa nucleofector 2, by Lonza (1 mentions)
Hcc1806, by American Type Culture Collection (1 mentions)
Htb 132, by American Type Culture Collection (1 mentions)
Crl 2336, by American Type Culture Collection (1 mentions)
Crl 2315, by American Type Culture Collection (1 mentions)
43 protocols using ccl 243
1
Umbilical Cord and Adult Blood Samples
2
Cultivation of A549 and K562 Cell Lines
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Lung adherent epithelial carcinoma A549 cells were obtained from ATCC (ATCC®; CCL-185). A549 cell lines were cultured in DMEM (Gibco, ThermoFisher) with 10% FBS and 1% penicillin/streptomycin (Gibco), and maintained until passage 20 before being discarded. Chronic myelogenous leukemia K562 cells were obtained from ATCC (ATCC®; CCL-243). K562 cell lines were cultured in RPMI-1640 (Life Technologies, ThermoFisher) with 10% FBS and 1% penicillin/streptomycin (Gibco), and maintained until passage 20 before being discarded. Human rhIL-15 and human rhIL-12 were from Gold Biotechnology, human rhIL-2 was from Akron Biotechnology. TGF-β was from ThermoFisher. All other reagents were from Sigma-Aldrich unless otherwise stated.
3
DENV2 and ZIKV Propagation in Cell Lines
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The DENV2 New Guinea reference strain and ZIKV (KU922960.1) were propagated in C3/36 cells derived from Aedes albopictus. These cells were grown in minimum essential medium (pH 7.2) supplemented with 10% fetal bovine serum (FBS), 1 mM sodium pyruvate, 2 mM l -glutamine, and 1X nonessential amino acids (NEAA) (all from FBS; Gibco, Carlsbad, CA, USA) and incubated at 34°C. DENV2 and ZIKV stocks were prepared by infecting a monolayer of C6/36 cells with 80–85% confluence for 24–48 h until a cytopathic effect was observed. The supernatant was collected, cleared of cells by centrifugation at 1,000 ×g for 10 min at 4°C, and concentrated in 0.22 μm Amicon columns to 1/10 of the starting volume. Next, 1/10 volume of sucrose-phosphate-glutamate stabilizer (2.18 mM sucrose, 38 mM monobasic K2HPO4, 72 mM dibasic K2HPO4, and 60 mM l -glutamic acid) was added, and the stocks were aliquoted and stored at −80°C until further use.
Vero and K562 (CCL-81 and CCL-243, respectively; ATCC) cell lines were cultured in RPMI medium (pH 7.8; Gibco) supplemented with 5% FBS, 2 mMl -glutamine, 1X NEAA, and 1X vitamins and incubated in a CO2 atmosphere at 37°C.
Drosophila melanogaster Schneider 2 (S2) cells (Gibco) were grown in Drosophila Schneider 2 culture medium supplemented with 10% FBS and 2 mMl -glutamine and incubated at 28°C in the absence of CO2.
Vero and K562 (CCL-81 and CCL-243, respectively; ATCC) cell lines were cultured in RPMI medium (pH 7.8; Gibco) supplemented with 5% FBS, 2 mM
Drosophila melanogaster Schneider 2 (S2) cells (Gibco) were grown in Drosophila Schneider 2 culture medium supplemented with 10% FBS and 2 mM
4
K562 Cell Culture and Transfection
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Virus Quantification and ADE Assays
6
Cell Culture of TNBC and K562 Lineages
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TNBC cell lines HCC-1806 (ATCC, CRL 2335™), HCC-1937 (ATCC, CRL-2336™), HCC-70 (ATCC, CRL-2315™), and MDA-MB-468 (ATCC, HTB-132™), and the chronic myelogenous leukemia-derived K562 cell line (ATCC, CCL-243™) were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). The cell lines were authenticated by DNA profiling using short tandem repeat (STR) analysis on an AmpFlSTR® Identifier™ PCR Amplification System at the National Institute of Genomic Medicine (INMEGEN), Mexico City, Mexico. The cells were used between passage 3 and passage 15. All the cells were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium (Biowest SAS, Nuaillé, France) that was supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Biowest SAS, Nuaillé, France), 100 U/mL penicillin, and 100 mg/mL streptomycin (Invitrogen, Carlsbad, CA, USA). The cells were maintained in a 5% CO2 humidified atmosphere at 37 °C.
7
Natural and Antibody-Dependent NK Cell Activation
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To measure general NK cell activation in vitro through natural cytotoxicity receptors (NCR), class I human histocompatibility-linked antigen (HLA) deficient erythroleukemia K562 cells (ATCC® #CCL 243™) were incubated with PBMC at an effector to target (E:T) ratio of 5:1 (2 × 106 PBMC:4 × 105 K562 cells). To selectively measure antibody-dependent NK cell activation in vitro, 2 µg anti-CD16 monoclonal antibody (LEAF™ 3G8, BioLegend), was added to 2 × 106 PBMC in 2 mL lymphocyte medium. After one hour, Brefeldin A (Sigma-Aldrich) was added to both stimulations to a final concentration of 10 µg/mL followed by an additional 15 h incubation whereby IFN-γ and TNF-α production was measured after intracellular staining. All cell lines used were cultured at 37 °C in 5% CO2 in lymphocyte medium and maintained in a log phase growth for labeling.
8
Antiproliferative Activity of Enzymosomes and Liposomes
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The antiproliferative activity of the formulations (enzymosomes and liposomes) was studied in K562 (myelogenous leukemia cell line, immortalized cell line K562 derived from a CML patient was obtained from ATCC, reference no. CCL-243 (Manassas, VA, USA)), Jurkat (T lymphocyte cell line, cells (clone E6-1) were obtained from ATCC), and H69 (small cell lung cancer, human classical SCLC cell line NCI-H69 was obtained from ATCC, reference no. HTB-119) cell lines. Free S. cerevisiae asparaginase expressed in P. pastoris (ASP), commercial E. coli asparaginase (Aginasa®, Kyowa Hakko Kirin Co., Ltd., Tokyo, Japan), and saline solution were used as controls.
The assays were performed according to the MTS protocol [30 (link)]. Briefly, cells that were maintained in the exponential growth phase were seeded in 96-well plates, at 2.5 × 103 cells per well. Afterward, cells were treated with ASP-liposomes, ASP-enzymosomes, or controls (0.08 to 10 IU/mL) and incubated at 37 °C and 5% CO2. After 48 h or 72 h, the MTS reagent (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) was added, and the plates were incubated at 37 °C for 2 h, after which the absorbances were read at 490 nm and 630 nm wavelength.
The assays were performed according to the MTS protocol [30 (link)]. Briefly, cells that were maintained in the exponential growth phase were seeded in 96-well plates, at 2.5 × 103 cells per well. Afterward, cells were treated with ASP-liposomes, ASP-enzymosomes, or controls (0.08 to 10 IU/mL) and incubated at 37 °C and 5% CO2. After 48 h or 72 h, the MTS reagent (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) was added, and the plates were incubated at 37 °C for 2 h, after which the absorbances were read at 490 nm and 630 nm wavelength.
9
Cloning of H2AFY1 Isoforms
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cDNA sequences of H2AFY1.1 and 1.2 were obtained from Addgene [H2AFY1.1 #45166, H2AFY1.2 # 45168, (Chadwick and Willard, 2001 (link))] and were cloned into a lenti-MND-IRES-GFP-Flag vector as described previously (Okeyo-Owuor et al., 2015 (link)). Lentivirus was generated in 293T cells (ATCC, CRL-3216) with the packaging plasmids pMD-G, pMD-Lg and REV. Virus was tittered using K562 cells (ATCC, CCL243).
10
Cell Culture Protocol for Cancer Research
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