All of the pediatric AML patient samples were obtained at the time of diagnosis from the University-Hospital of Padua and stratified according to the AIEOP AML 2002/01 protocol AML 2002/01 [47 (link)]. Patient characteristics are listed in Supplementary Table 1 . Seven BM samples from healthy donors were obtained as control. All human myeloid cell lines (THP-1, NOMO-1, OCI-AML4, ML2, HL60, K562, Kasumi-1, NB4, U-937, SEM and REH) were obtained from DSMZ (Braunschweig, Germany) and 293T cells were obtained from ATCC (Manassas, VA, USA). All cell lines were maintained under standard conditions suggested by the manufacturer.
Thp 1
Manufactured by Leibniz Institute DSMZ
Sourced in Germany
The THP-1 is a human acute monocytic leukemia cell line. It is a widely used in vitro model for studying monocyte and macrophage biology.
Automatically generated - may contain errors
Lab products found in correlation
Mv4 11, by Leibniz Institute DSMZ (12 mentions)
Hl 60, by Leibniz Institute DSMZ (11 mentions)
Molm 13, by Leibniz Institute DSMZ (10 mentions)
Oci aml3, by Leibniz Institute DSMZ (7 mentions)
Oci aml2, by Leibniz Institute DSMZ (6 mentions)
Nomo 1, by Leibniz Institute DSMZ (5 mentions)
Kasumi 1, by Leibniz Institute DSMZ (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Kg 1a, by Leibniz Institute DSMZ (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Skm 1, by Leibniz Institute DSMZ (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Monomac6, by Leibniz Institute DSMZ (3 mentions)
Ultraglutamine 20mm solution, by Lonza (2 mentions)
Human interleukin 4 il 4, by Miltenyi Biotec (2 mentions)
Rneasy mini kit, by Qiagen (2 mentions)
Rpmi 1640, by Lonza (2 mentions)
Mycoalert kit, by Lonza (2 mentions)
Ht 29, by Leibniz Institute DSMZ (2 mentions)
Molm 14, by Leibniz Institute DSMZ (2 mentions)
41 protocols using thp 1
1
Pediatric AML Diagnostic Samples
2
Functional Profiling of Antigen-Specific CD8+ T Cells
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Dextramer‐selected P1‐specific CD8+ T cells were co‐cultured with the HLA‐A*02 AML cell line THP1 (DSMZ, GE, EU) in a ratio 10:1. T2 cells loaded with P1 or the irrelevant HERV‐derived peptide (SMDDQLNQL) were used as positive and negative controls of specificity, respectively. After 1 h of co‐culture, Golgi Plug (BD, NJ, US) and CD107a antibody (BD, NJ, US) were added into wells and co‐cultures were maintained for further 4 h. An anti‐human MHC I blocking Ab (Clone W6/32, InVivoMab, BioXcell, NH, US) was added in selected wells for 1 h before co‐culture (50 μg/ml) and maintained at 10 μg/ml during the entire time of the co‐culture to neutralize the HLA‐A*02 dependent T cell activation, as control. Viability (Zombie NearIR, Biolegend, CA, US), extracellular (CD3, BV421/ CD8, FITC both Biolegend, CA, US), intracellular staining (IFN‐γ PE, Biolegend, CA, US and TNF‐α Pe‐Cy7, BD, NJ, US) and fixation were then performed and samples were analyzed on a LSR Fortessa (BD, NJ, US).
3
Monocytic leukemia cell line THP-1 protocol
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The human monocytic leukemia cell line THP-1 was purchased from DSMZ (Germany). Growth media (RPMI 1640) and Ultraglutamine 20 mM solution were purchased from Lonza (Belgium). Foetal bovine serum (FBS), phorbol 12-myristate 13-acetate (PMA), pure ethanol and ultra-pure water (W3500) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Human Interleukin 4 (IL-4) was purchased from Macs-Miltenyi Biotec (Germany). Cell proliferation reagent WST-1 was purchased from Roche Diagnostics GmbH (Germany) and the dye reagent for the Bradford protein assay was purchased from Bio-Rad Laboratories GmbH (Germany). RNeasy Mini kit, RT first strand kit and RT SYBR Green Rox Master Mix were from Qiagen, (CA, USA). The 384 RT-Array custom was made by Sabiosciences (Qiagen Company, CA, USA). Arnica m. was produced by Boiron Laboratoires (Lyon, France) according to the French Homeopathic pharmacopoeia and provided as a first centesimal dilution of the whole hydroalcoholic extract (Mother Tincture, MT) in 30% ethanol/distilled water. The MT was characterised by a content of sesquiterpene lactones of 0.036% (W/V).
4
THP-1 Cell Culture Protocol
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The in vitro experiments were performed using the THP-1 (human acute monocytic leukemia) cell line (DSMZ GmbH, Braunschweig, Germany), cultured in RPMI complete medium [90% RPMI-1640 (PAA GmbH, Cölbe, Germany), 10% fetal bovine serum (FBS, PAA GmbH); 100 U/ml penicillin; 0.1 mg/ml streptomycin (PAA GmbH)].
5
Culturing Acute Myeloid Leukemia Cell Lines
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Eight human acute myeloid leukemia cell lines (KG-1a, KG-1, HL-60, NB-4, ML-2, THP-1, MV-4-11, U-937) covering M0 to M5 FAB stages were purchased from DSMZ (German Collection of Microorganisms & Cell Cultures, Braunschweig, Germany), used at a maximum of 20 passages, and frequently tested for the absence of mycoplasma contamination. Cells were cultured in RPMI media, supplemented with 10% fetal bovine serum (FBS), at 37 °C in fully humidified air and 5% CO2. Cells were harvested from the culture at the exponential growth phase.
6
Evaluating CAR-DOT Cytotoxicity and Cytokine Production
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The cell lines MOLM13, THP-1 and Jurkat were purchased from DSMZ (Germany) and expanded according to DSMZ recommendations. Target cells (cell lines and primary AML blasts) were labeled with 3 µM eFluor 670 (eBioscience) and incubated with CAR-DOTs or mock-DOTs at different Effector:Target (E:T) ratios for the indicated time periods in complete DOT media. CAR-DOT-mediated cytotoxicity was determined by analyzing the residual alive (7-AAD–) eFluor 670+ target cells at each time point and E:T ratio. Absolute cell counts were determined using Trucount absolute count beads (BD Biosciences).28 (link) Cytokine production was measured by G-Series Human Cytokine Antibody Array 4000 from RayBiotech in supernatants harvested after 48 hours-incubation CAR-DOT/mock-DOT (four donors) with two primary AML samples.
7
Establishment of Resistant AML Cell Lines
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The AML cell line THP-1 was obtained from DSMZ (Braunschweig, Germany). Cytarabine (AraC) and doxorubicin (DOX)-resistant sublines were established by continuous exposure to stepwise increasing drug concentrations as previously described [19] and derived from the Resistant Cancer Cell Line (RCCL) collection (www.kent.ac.uk/stms/cmp/RCCL/RCCLabout.html) [20] . There were no clonal isolation processes involved, thus each subline is expected to be a mixture of clones able to grow optimally under the drug exposure. To ensure AraC and DOX resistance during the entirety of the study, IC50 concentrations against AraC and DOX were determined in the resistant and paired controls each 2–3 weeks. All the cell lines were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 4 mM glutamine and 1% penicillin and streptomycin at 37 °C in a humidified incubator with 5% CO2. Particularly, AraC and DOX resistant cells were maintained with 8 µM of AraC or 20 pM of DOX, respectively.
8
Diverse Cell Lines for Research
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Four cell lines were utilized in this work: A549 pulmonary carcinoma epithelial cell line (A549, DSMZ-ACC 107, Germany, RPMI 1640 supplemented with 10% fetal calf serum, FCS), HaCaT human keratinocyte cell line (HaCaT, CLS-300493, Germany, DMEM supplemented with 10% FCS), THP-1 human acute leukemia monocyte cell line (THP-1, DSMZ-ACC 16, Germany, RPMI 1640 supplemented with 10% FCS), and DC2.4 murine dendritic cell line (DC2.4, SCC142-Merck Millipore, Germany, RPMI 1640 supplemented with 10% FCS + 1x nonessential amino acids + 1x HEPES buffer + 0.0054x β-mercaptoethanol).
9
THP-1 Cell Culture Protocol
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The in vitro experiments were performed using the THP-1 (human acute monocytic leukemia) cell line (DSMZ GmbH, Braunschweig, Germany), cultured in 90% RPMI-1640 (PAA GmbH, Cölbe, Germany), 10% FBS (PAA GmbH); 100 U/mL penicillin; and 0.1 mg/mL streptomycin (PAA GmbH). All experiments were carried out in medium with 10% FBS.
10
Establishment and Maintenance of Myeloid Leukemia Cell Lines
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Acute monocytic leukemia cell lines THP-1 and MV4-11 were obtained from DSMZ and bone marrow stromal cell line HS-5 from ATCC. Cell culture reagents were obtained from Thermo Fisher Scientific unless otherwise stated.
Acute myeloid leukemia (THP-1 and MV4-11) were maintained in RPMI 1640 medium supplemented with 10% (v/v) heat-inactivated FBS, 2 mM L-glutamine, penicillin and streptomycin in a humidified incubator at 37°C and 5% CO2. Establishment of FFM05 cells has been previously described [25 (link)]. FFM05 cells were maintained in X-Vivo medium (Lonza) supplemented with 10% (v/v) FBS HyClone, 2 mM L-glutamine, and human recombinant growth factory: thrombopoietin (TPO) (25 ng/ml), stem cell factor (SCF) (50 ng/ml), FMS-related tyrosine kinase 3 ligand (FLT3-I) ligand (50 ng/ml), and interleukin-3 (IL-3) (20 ng/ml) (Miltenyi Biotech).
Acute myeloid leukemia (THP-1 and MV4-11) were maintained in RPMI 1640 medium supplemented with 10% (v/v) heat-inactivated FBS, 2 mM L-glutamine, penicillin and streptomycin in a humidified incubator at 37°C and 5% CO2. Establishment of FFM05 cells has been previously described [25 (link)]. FFM05 cells were maintained in X-Vivo medium (Lonza) supplemented with 10% (v/v) FBS HyClone, 2 mM L-glutamine, and human recombinant growth factory: thrombopoietin (TPO) (25 ng/ml), stem cell factor (SCF) (50 ng/ml), FMS-related tyrosine kinase 3 ligand (FLT3-I) ligand (50 ng/ml), and interleukin-3 (IL-3) (20 ng/ml) (Miltenyi Biotech).
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