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Hl 60 cells

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The HL-60 cells are a human promyelocytic leukemia cell line. They are a widely used model system for studying various aspects of myeloid cell differentiation and function.

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Lab products found in correlation

Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Hl 60 cells, by American Type Culture Collection (2 mentions) Fetal bovine serum (fbs), by Sartorius (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Lonza (1 mentions) U937 cells, by American Type Culture Collection (1 mentions) Kasumi 1 cells, by American Type Culture Collection (1 mentions) Rpmi 1640 growth medium, by Thermo Fisher Scientific (1 mentions) Thp 1 cell, by American Type Culture Collection (1 mentions) Iscove s modified dulbecco s medium, by Thermo Fisher Scientific (1 mentions) Hl 60 cells, by Shanghai Cell Bank (1 mentions) Thp 1 cells, by Shanghai Cell Bank (1 mentions) Tomatidine, by Merck Group (1 mentions) α tomatine, by Merck Group (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Cholesterol, by Merck Group (1 mentions)

9 protocols using hl 60 cells

1

Differentiation of HL-60 Cells into Neutrophils

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Human promyelocytic HL-60 cells (KCLB, South Korea) were grown in RPMI-1640 medium (Thermo Scientific) supplemented with 10% FBS (Lonza), 0.1 mM nonessential amino acids, 50 U/mL penicillin, and 50 μg/mL streptomycin at 37 °C in a humidified 5% CO2 incubator. The HL-60 cells were induced to undergo differentiation along the neutrophil lineage by seeding aliquots of HL-60 cell suspension (2 × 106 cells/mL) onto a tissue culture flask and growing for 5 days in the presence of 1.25% dimethyl sulfoxide (DMSO). Complete differentiation of HL-60 cells into neutrophil-like cells (abbreviated dHL-60) was confirmed by nuclear segmentation and granule formation, spectrophotometry at 540 nm, and flow cytometry of FITC-conjugated monoclonal antibodies against CD11b, CD16, and CD33 (BD Biosciences). Thereafter, dHL-60 cells were cryopreserved at a concentration of 2 × 106 cells/mL in FBS containing DMSO (10% v/v) until seeding into the EC channel.
Han S., Shin Y., Jeong H.E., Jeon J.S., Kamm R.D., Huh D., Sohn L.L, & Chung S. (2015). Constructive remodeling of a synthetic endothelial extracellular matrix. Scientific Reports, 5, 18290.
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2

Culturing Human Hematopoietic Cell Lines

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Kasumi-1 cells (ATCC CRL-2724), HL60 cells (ATCC CCL-240), THP-1 cells (ATCC TIB-202) and U937 cells (CRL-1593) were purchased from ATCC. Cells were cultured following the manufacturer’s culture method. Kasumi cells were cultured with RPMI-1640 growth medium (Gibco, 11875093) containing 20% FBS. HL-60 cells were cultured with IMDM containing 20% FBS (Thermofisher 12440053). THP-1 cells were cultured with RPMI-1640 containing 10% FBS. U937 cells were cultured with RPMI-1640 containing 10% FBS.
Zhang N., Long Y, & Devreotes P.N. (2001). Gγ in Dictyostelium: Its Role in Localization of Gβγ to the Membrane Is Required for Chemotaxis in Shallow Gradients. Molecular Biology of the Cell, 12(10), 3204-3213.
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3

Culturing Leukemia Cell Lines

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Cells and cell culture. The THP-1 cells (derived from a patient with monocytic leukemia) and HL-60 cells (derived from a patient with promyelocytic leukemia) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The K562 cells (derived from a patient with chronic myelogenous leukemia) were obtained from CHI Scientific (Jiangyin, China). The U937 cells (derived from a patient with histiocytic lymphoma) were purchased from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences (Beijing, China). The HL-60 cells were cultured in Iscove's modified Dulbecco's medium (Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Biological Industries, Beit Haemek, Israel), 100 U/ml penicillin and 0.1 mg/ml streptomycin. The THP-1, K562, and U937 cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Inc.) containing 10% FBS (Biological Industries), 100 U/ml penicillin and 0.1 mg/ml streptomycin. All cell lines were maintained in an incubator (Shanghai Lishen, Shanghai, China) with 5% CO 2 at 37˚C.
Sun L., Sun C., Sun J, & Yang W. (2019). Downregulation of ENDOCAN in myeloid leukemia cells inhibits proliferation and promotes apoptosis by suppressing nuclear factor‑κB activity. Molecular medicine reports, 19(4).
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4

HL-60 Cell Culture Protocol

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HL-60 cells were obtained from the American Type Culture Collection (Rockville, MD, USA). Tomatidine, α-tomatine and cholesterol were purchased from Sigma-Aldrich (St. Louis, MO, USA). RPMI-1640, penicillin-streptomycin, L-glutamine and fetal bovine serum (FBS) were purchased from Gibco-BRL (Grand Island, NY, USA). HL-60 cells were maintained in RPMI-1640 culture medium containing 10% FBS that was supplemented with penicillin (100 U/ml), streptomycin (100 μg/ml) and L-glutamine (300 μg/ml) (Gibco-BRL). Cultured cells were grown at 37°C in a humidified atmosphere of 5% CO2 and were passaged twice a week.
HUANG H., CHEN S., VAN DOREN J., LI D., FARICHON C., HE Y., ZHANG Q., ZHANG K., CONNEY A.H., GOODIN S., DU Z, & ZHENG X. (2015). α-tomatine inhibits growth and induces apoptosis in HL-60 human myeloid leukemia cells. Molecular Medicine Reports, 11(6), 4573-4578.
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5

Generating Etoposide-Resistant HL60 Cell Lines

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HL60 cells (CLS GmBH, Germany) were cultured in RPMI media (Gibco, Life Technologies, USA) supplemented with 10% FBS (Gibco) and maintained at 37°C in a humidified incubator. Etoposide resistant cells HL60-EtopRwere generated by culturing HL60 cells in
gradually incremental doses of Etoposide (2nM to 1µM) over a period of 2 months.Single cell clones of etoposide resistant cells were isolated by culturing the drug selected cells in methylcellulose-based semisolid media and picking up isolated colonies
of resistant cells.These isolated clones were named: HL60-EtopRH1A, HL60-EtopRH1B, and HL60-EtopRH1C.
Alghamdi R.H., Ahmed F., Ibrahim S.M., Pushparaj P.N., Schulten H.J., Abuzenadah A.M, & Almalki A.L. (2022). Molecular determinants of etoposide resistance in HL60 cells. Bioinformation, 18(10), 894-899.
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6

Culturing Human Leukemia Cell Lines

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Human HL60 cell line and U937 cell line were purchased from Shanghai Micro Mongolian Life Science Co., Ltd (Shanghai, People’s Republic of China). HL60 cells and U937 cells were cultured in RPMI-1640 medium with 10% fetal bovine serum (FBS) (Gibco BRL, Rockville, MD, USA), 100 U/mL penicillin G and 100 μg/mL streptomycin at 37°C in a humidified 5% CO2 atmosphere. The logarithmic growth phase cells were used for further experiments.
Ye Q., Liao X., Fu P., Dou J., Chen K, & Jiang H. (2016). Portulacerebroside A inhibits adhesion, migration, and invasion of human leukemia HL60 cells and U937 cells through the regulation of p38/JNK signaling pathway. OncoTargets and therapy, 9, 6953-6963.
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7

Cell Line Treatments and Responses

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Human acute promyelocytic leukemia HL-60 cells and human lung carcinoma A549 cells were obtained from ATCC (Manassas, VA). The HL-60 cells were cultured in Isocove’s modified Dulbecco’s medium (IMDM, Gibco) supplemented with 10% v/v heat-inactivated fetal bovine serum (Atlanta Biologicals) at 37 °C with 5% CO2 atmosphere and treated with 225 μM Zn or deionized water. The A549 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) with 10% v/v heat-inactivated fetal bovine serum (Atlanta Biologicals) at 37 °C with 5% CO2 atmosphere and treated with either 10 pM of the Clostridium difficile toxin TcdB23 (link) or 20 mM Hepes/50 mM NaCl buffer.
Gutierrez D.B., Gant-Branum R.L., Romer C.E., Farrow M.A., Allen J.L., Dahal N., Nei Y.W., Codreanu S.G., Jordan A.T., Palmer L.D., Sherrod S.D., McLean J.A., Skaar E.P., Norris J.L, & Caprioli R.M. (2018). An Integrated, High-Throughput Strategy for Multiomic Systems Level Analysis. Journal of proteome research, 17(10), 3396-3408.
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8

Differentiating HL-60 Cells into Neutrophils

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HL-60 cells were obtained from the Bioresource Collection and Research Center. The HL-60 cells were maintained in RPMI 1640 medium with L-glutamine (GeneDireX, Inc.) supplemented with 20% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and antibiotics, including 100 U/ml penicillin, 100 µg/ml streptomycin and 0.25 µg/ml amphotericin B, at 37˚C in a humidified atmosphere with 5% CO2. The cell density was maintained between 1x105 and 1x106 cells/ml. To differentiate the HL-60 cells into a neutrophil-like phenotype, the cells were cultured at a density of 1x106 cells/ml in RPMI 1640 medium containing L-glutamine supplemented with 10% fetal bovine serum, 10 mM HEPES (Gibco; Thermo Fisher Scientific, Inc.), the aforementioned antibiotics and 1.25% DMSO for 6 days.
Liu K.T., Yeh I.J., Hsu Y.L, & Yen M.C. (2024). Transcriptomic analysis of lipoteichoic acid‑treated undifferentiated and neutrophil‑like differentiated HL‑60 cells. Experimental and Therapeutic Medicine, 27(4), 158.
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9

Differentiation of HL-60 cells into Neutrophils

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2.1. Culture and differentiation of HL-60 cells HL-60 cells (ATCC) were cultured in RPMI-1640 (Gibco), supplemented with 2 mM GlutaMax (Gibco), 10% heatinactivated bovine growth serum (Hyclone), 100 U/ml penicillin (Gibco), 100 μg/ml streptomycin (Gibco) and 0.25 μg/ml amphotericin B (Sigma). To differentiate HL-60 cells into neutrophils, 1.25% of DMSO (Sigma-Aldrich) was added to the complete medium for 5 days with medium replenishment every 2 days.
Jennings S., Ng H.P, & Wang G. (2019). Establishment of a ΔF508-CF promyelocytic cell line for cystic fibrosis research and drug screening. Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society, 18(1).
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