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Hl 60

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The HL-60 is a cell line derived from a patient with acute promyelocytic leukemia. It is a widely used model for studying the differentiation and function of myeloid cells.

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Lab products found in correlation

Hl 60, by American Type Culture Collection (15 mentions) Thp 1, by American Type Culture Collection (7 mentions) Rpmi 1640 medium, by Merck Group (7 mentions) Streptomycin, by Merck Group (7 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Penicillin, by Merck Group (6 mentions) Thp 1, by Merck Group (5 mentions) Fetal bovine serum (fbs), by Merck Group (5 mentions) Dimethyl sulfoxide (dmso), by Merck Group (4 mentions) Kasumi 1, by Merck Group (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Rpmi 1640, by Merck Group (4 mentions) Phorbol myristate acetate (pma), by Merck Group (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) L glutamine, by Thermo Fisher Scientific (3 mentions) Mycoalert mycoplasma detection kit, by Lonza (3 mentions) Kasumi 1, by American Type Culture Collection (3 mentions) Mv4 11, by American Type Culture Collection (3 mentions) Oci aml3, by Leibniz Institute DSMZ (3 mentions) Molm 13, by Merck Group (3 mentions)

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HL-60 cells (5 citations)

38 protocols using hl 60

1

Characterization of Drug-Resistant Cancer Cells

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The following human
cancer cell lines were used in this study: the colon carcinoma-derived
cell line SW480 and lung fibroblasts WI-38 (obtained from the American
Tissue Culture Collection), the ovarian carcinoma-derived cell line
A2780 (obtained from Sigma-Aldrich), the acute promyelocytic leukemia-derived
cell line HL-60 (obtained from Dr. M. Center, Kansas State University),
the cervix carcinoma-derived cell line KB-3-1 and the colchicine resistant
subline KBC-1 (obtained from Dr. D. W. Shen, Bethesda, Maryland).
SW480 and WI-38 cells were grown in MEM with 10% FCS and A2780, KB-3-1,
KBC-1, and HL-60 cells were cultured in RPMI 1640 supplemented with
10% FCS. SW480/Tria cells were generated by continuous exposure of
SW480 cells to increasing concentrations of Triapine (starting point,
0.05 μM; end point, 20 μM) over a period of one year.19 (link) Triapine was administered to the cells once
every other week at the day after passage, when cells had attached
to the culture flasks.
Synergism is expressed by the combination
index (CI) according to Chou and Talalay49 (link) using CalcuSyn software (Biosoft, Ferguson, MO, USA). CI < 0.9,
CI = 0.9–1.2, or CI > 1.2 represent synergism, additive
effects,
and antagonism, respectively.
Kowol C.R., Miklos W., Pfaff S., Hager S., Kallus S., Pelivan K., Kubanik M., Enyedy É.A., Berger W., Heffeter P, & Keppler B.K. (2016). Impact of Stepwise NH2-Methylation of Triapine on the Physicochemical Properties, Anticancer Activity, and Resistance Circumvention. Journal of Medicinal Chemistry, 59(14), 6739-6752.
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2

Differentiation of Monocytic and Granulocytic Cell Lines

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The human monocytic cell line THP-1, and the promyelocytic cell line NB4 and HL-60 were purchased from ATCC (Manassas, USA) and maintained in RPMI1640 supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA). Cells were screened by PCR for authentication and by immunofluorescence tests for freedom from mycoplasma contamination. Monocytic differentiation of THP-1 and HL-60 cells was induced by treating the cells with 50 nM PMA (Sigma-Aldrich, Deisenhofen, Germany) for 0, 24, 48, and 72 h. Granulocytic differentiation of NB4 and HL-60 cells was induced by treating the cells with 1 μM ATRA (Sigma-Aldrich, Deisenhofen, Germany). No cell lines used in this study were found in the database of commonly misidentified cell lines that is maintained by ICLAC and NCBI Biosample. Cell lines were tested for mycoplasma contamination but have not been re-authenticated.
Zhao H., Wang X., Yi P., Si Y., Tan P., He J., Yu S., Ren Y., Ma Y., Zhang J., Wang D., Wang F, & Yu J. (2017). KSRP specifies monocytic and granulocytic differentiation through regulating miR-129 biogenesis and RUNX1 expression. Nature Communications, 8, 1428.
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3

Establishment of ADR-resistant Leukemia Cell Lines

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Human acute leukemia cell lines (HL60 and THP-1) obtained from ATCC (Manassas, VA, USA) were maintained in Iscove's Modified Dulbecco's Medium (IMDM, Gibco, Invitrogen Ltd, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Cripion, Industria Brasileira, Andradina, SP, Brazil), 1% (v/v) antibiotics (100 U ml−1 penicillin and 100 μg ml−1 streptomycin, Gibco) at 37 °C in a humidified 5% CO2 incubator. ADR-resistant AML cell lines (HL60/ADR and THP-1/ADR) were established as described previously18 (link) as follows: (1) HL60 and THP-1 cells in the logarithmic phase were treated with 0.8 μM of ADR (Sigma-Aldrich, St. Louis, MO, USA) for 12 h, and then fresh ADR-free medium was replaced; (2) cells were allowed to reach the logarithmic phase before another ADR treatment cycle, until the treatment showed no significant influence on cell morphology and proliferation. Additional 0.5 μM of ADR was added into the culture medium to maintain the ADR-resistant phenotype of HL60/ADR and THP-1/ADR.
Li Q, & Wang J. (2019). Long noncoding RNA ZFAS1 enhances adriamycin resistance in pediatric acute myeloid leukemia through the miR-195/Myb axis. RSC Advances, 9(48), 28126-28134.
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4

Differentiation of HL-60 Cells into Neutrophils

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The human promyelocytic leukemia cell line (HL-60) was purchased from iCell Bioscience Inc. (Shanghai, China). HL-60 cells were induced to differentiate into neutrophil-like cells (HL-60N) by culturing them in RPMI-1640 medium supplemented with 10% FBS and 1.25% DMSO (Sigma-Aldrich) for 4 days (46 (link)). For the transfection experiment, HL-60N cells were seeded at a density of 5 × 105 cells/well in plates and incubated for 48 h in the presence of 100 nM TOB1 small interfering RNA (si- TOB1) or negative control siRNA (si-NC) obtained from General Biol (Chuzhou, China) and the Lipofectamine 2000 transfection reagent (Invitrogen, Carlsbad, CA, USA). All cells were maintained in a humidified incubator at 37°C with 5% CO2. The sequences of si- TOB1 and si-NC are provided in Table S1.
Zhang J., Li Y., Chen J., Huang T., Lin J., Pi Y., Hao H., Wang D., Liang X., Fu S, & Yu J. (2024). TOB1 modulates neutrophil phenotypes to influence gastric cancer progression and immunotherapy efficacy. Frontiers in Immunology, 15, 1369087.
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5

Propagation and Maintenance of Primary T-cells and Human Leukemia Cell Lines

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The isolated CD3, CD4, and CD8 primary T-cell and human leukemia K562, HL60 and KG1 cell lines (purchased from Sigma) were propagated in culture media: RPMI 1640 with 10% heat-inactivated fetal calf serum (FBS; Thermo Fisher Scientific, Merelbeke, Belgium) and with L-glutamine, penicillin and streptomycin at standard concentrations [all from Gibco, Invitrogen, United Kingdom (UK)] in humidified air with 5% CO2 at 37 °C. The cell culture media were renewed every 2–3 days. Once the cells were at or near confluence, they were subcultured. Melanoma primary cultures were regularly assessed for mycoplasma contamination via a MycoAlert® Mycoplasma Detection Kit (Lonza, Rockland, ME, USA).
Otmani K., Rouas R., Lagneaux L., Krayem M., Duvillier H., Berehab M, & Lewalle P. (2023). Acute myeloid leukemia-derived exosomes deliver miR-24-3p to hinder the T-cell immune response through DENN/MADD targeting in the NF-κB signaling pathways. Cell Communication and Signaling : CCS, 21, 253.
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6

Cytotoxicity Evaluation of DC-S285 in Cancer Cell Lines

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MCF7, HL60, MV4-11, K562, Kasumi-1, U937, THP1, and Jurkat cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). Fetal bovine serum was purchased from Life Technologies. MCF7 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum at 37 °C in an incubator with 5% CO2 atmosphere. HL60 and MV4-11 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum. K562, Kasumi-1 and HL60 were cultured in 96 plate at 10,000/well for 2 h. DC-S285 was dissolved in DMSO (Sigma) and then stored at 4 °C. The cells were incubated with DC-S285 at different concentrations ranging from 0 to 100 µM for approximately 72 h. The activities of DC-S239 against MCF7, HL60, MV4-11, K562, Kasumi-1, U937, THP1, and Jurkat were measured by the alamarBlue assays and MTT assays.
Ding H., Lu W.C., Hu J.C., Liu Y.C., Zhang C.H., Lian F.L., Zhang N.X., Meng F.W., Luo C, & Chen K.X. (2018). Identification and Characterizations of Novel, Selective Histone Methyltransferase SET7 Inhibitors by Scaffold Hopping- and 2D-Molecular Fingerprint-Based Similarity Search. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(3), 567.
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7

Characterization of AML Cell Lines

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Acute Myeloid Leukemia (AML) cell lines HL-60, MOLM-13, and OCI-AML3 were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig, Germany). Cell lines were maintained at 37 °C and 5% CO2 in appropriate media (HL60 and MOLM13 in RPMI medium (Sigma-Aldrich, Darmstadt, Germany) containing 10% heat-inactivated FBS (Biochrom, Berlin, Germany) and OCIAML3 in alpha-MEM medium (Sigma-Aldrich, Darmstadt, Germany) containing 20% heat-inactivated FBS). Cell lines were routinely verified for mycoplasma contamination using Venor®GeM Mycoplasma Detection Kit (Sigma-Aldrich, Darmstadt, Germany). Cell lines were authenticated by Multiplexion, Germany. The inhibitors cytarabine, daunorubicin, and vorinostat were purchased from Selleckchem (Houston, TX, USA). The MEK inhibitor PD98059 was purchased from Merck, Darmstadt, Germany.
More P., Ngaffo J.A., Goedtel-Armbrust U., Hähnel P.S., Hartwig U.F., Kindler T, & Wojnowski L. (2023). Transcriptional Response to Standard AML Drugs Identifies Synergistic Combinations. International Journal of Molecular Sciences, 24(16), 12926.
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8

Apoptosis Assessment of T Lymphocytes and Leukemia Cells

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The isolated T lymphocytes and human leukemia K562, HL60 and KG1 cells (purchased from Sigma) were maintained in medium containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Merelbeke, Belgium) and cultured at 37 °C in humidified air containing 5% CO2. T lymphocyte cell death was assessed by an annexin-V-FITC/propidium iodide (PI) and annexin-V-APC/7-AAD-based apoptosis detection kit from BD Biosciences according to the manufacturer’s instructions. Cells were seeded in 12-well plates in the presence of 5 μg/mL phytohemagglutinin (PHA-L, Sigma-Aldrich) and 20 U/mL IL-2 (from Sigma). After six days, the cells were harvested, washed twice with PBS-EDTA, stained with annexin-V/PI and analyzed in a FACS machine (NAVIOS-Beckman Coulter, Suarlée, Belgium), and the data generated were analyzed by KALUZA software (Beckman Coulter).
Moussa Agha D., Rouas R., Najar M., Bouhtit F., Fayyad-Kazan H., Lagneaux L., Bron D., Meuleman N., Lewalle P, & Merimi M. (2020). Impact of Bone Marrow miR-21 Expression on Acute Myeloid Leukemia T Lymphocyte Fragility and Dysfunction. Cells, 9(9), 2053.
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9

Culturing Leukemia Cell Lines

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NB4, a human APL cell line with t(15;17), was obtained from the Deutsche Sammalung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany). A human promonocytic leukemia cell line, U-937, and a human acute lymphoblastoid leukemia cell line, MOLT-4, were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). A human myeloid leukemia cell line, HL-60, was obtained from the RIKEN cell bank (Ibaraki, Japan). All cell lines were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS (Sigma for NB4 and HL-60; Nichirei for U-937 and MOLT-4), 100 U/mL of penicillin and 100 µg/mL of streptomycin in a humidified 5% CO2 atmosphere at 37 °C.
Yuan B., Kikuchi H., Li J., Kawabata A., Yao K., Takagi N, & Okazaki M. (2023). Cytotoxic Effects of Darinaparsin, a Novel Organic Arsenical, against Human Leukemia Cells. International Journal of Molecular Sciences, 24(3), 2282.
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10

Cell Line Cultivation and Treatment

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HL-60, KG-1a, K562, HL-60/ADR,
KG-1a/ADR, K562/ADR, SW1990, and SH-SY5Y were purchased from ATCC
and authenticated by short tandem repeat testing. All the cell lines
were maintained in appropriate medium as the manufacturer suggested.
HL-60, KG-1a, K562, HL-60/ADR, KG-1a/ADR, and K562/ADR were cultured
in RPMI 1640 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented
with 12% fetal bovine serum (FBS; Gibco BRL, Gaithersburg, ML, USA),
penicillin (100 IU/mL), and streptomycin (100 μg/mL) in a humidified
incubator at 37 °C and 5% CO2/95% air. Prior to each
experiment, HL-60/ADR, KG-1a/ADR, and K562/ADR cells were treated
with 1 μmol/mL ADR (Sigma-Aldrich, St. Louis, MO, USA) for 10–28
days and then cultured for 10 days without ADR exposure. SW1990 and
SH-SY5Y were cultured in DMEM medium (Sigma-Aldrich, St. Louis, MO,
USA) supplemented with 10% fetal bovine serum (FBS; Gibco BRL, Gaithersburg,
ML, USA), penicillin (100 IU/mL), and streptomycin (100 μg/mL).
All the cell lines were tested for mycoplasma contamination and were
found to be negative.
For experiments, cells were treated with
compounds for different concentrations with 0.1% dimethyl sulfoxide
(DMSO; Sigma-Aldrich, St. Louis, MO, USA) as solvent control. Where
indicated, 100 μM pervanadate was added 10 min before cell lysis.
Li J., Li S., Guo J., Li Q., Long J., Ma C., Ding Y., Yan C., Li L., Wu Z., Zhu H., Li K.K., Wen L., Zhang Q., Xue Q., Zhao C., Liu N., Ivanov I., Luo M., Xi R., Long H., Wang P.G, & Chen Y. (2018). Natural Product Micheliolide (MCL) Irreversibly Activates Pyruvate Kinase M2 and Suppresses Leukemia. Journal of Medicinal Chemistry, 61(9), 4155-4164.
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