The human leukemia cell lines, K562 (chronic myeloid leukemia cells), HL-60 (acute myeloid leukemia cells), MV-4-11 (biphenotypic B myelomonocytic leukemia cells), Jurkat (T-cell acute lymphoblastic leukemia cells), and K562/A02 (multidrug resistance K562) were from the American Type Culture Collection; HL-60/ADR (multidrug resistance HL-60) was from the Institute of Hematology & Blood Diseases Hospital of Chinese Academy of Medical Sciences & Peking Union Medical College. Cells were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS and 2 mM glutamine in a humidified incubator with 5% CO2 and 95% air.
Hl 60
Manufactured by American Type Culture Collection
Sourced in United States, Germany, United Kingdom, China, Australia
The HL-60 is a human promyelocytic leukemia cell line. It is a well-established in vitro model system for studying cellular differentiation and hematopoiesis.
Automatically generated - may contain errors
Lab products found in correlation
Thp 1, by American Type Culture Collection (152 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (97 mentions)
Mv4 11, by American Type Culture Collection (69 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (62 mentions)
Kasumi 1, by American Type Culture Collection (50 mentions)
Molm 13, by American Type Culture Collection (36 mentions)
Jurkat, by American Type Culture Collection (33 mentions)
Mcf 7, by American Type Culture Collection (32 mentions)
Rpmi 1640, by Thermo Fisher Scientific (28 mentions)
Penicillin, by Thermo Fisher Scientific (27 mentions)
Streptomycin, by Thermo Fisher Scientific (27 mentions)
Hl 60, by Thermo Fisher Scientific (27 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (26 mentions)
Hek293t, by American Type Culture Collection (25 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (24 mentions)
Hepg2, by American Type Culture Collection (23 mentions)
Hct116, by American Type Culture Collection (20 mentions)
Mda mb 231, by American Type Culture Collection (19 mentions)
Oci aml3, by American Type Culture Collection (18 mentions)
Molt 4, by American Type Culture Collection (17 mentions)
480 protocols using hl 60
1
Culturing Human Leukemia Cell Lines
2
Leukemia Cell Line Cultivation and Proliferation
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Human leukemia cell lines Jurkat, HL-60, and K-562 were obtained from ATCC. The normal bone marrow cell line (HS-5) were purchased from the American Type Culture Collection. The bone marrow cell line and leukemia cells were cultured in alpha-minimal essential medium (ThermoFisher). HEK293T cells were cultured in Rosewell Park Memorial Institute 1640 (ThermoFisher). All culture medium was supplied with 10% fetal bovine serum (ThermoFisher), 100 units/ml penicillin and streptomycin (Gibco). The cells were cultured at 37 °C and 5% CO2. To analyze the proliferation rate of the cells, cells were seeded at 1 × 104 or 1 × 103 cells/ml in 10-cm dishes and the cell number was counted every day.
3
Expansion and Transduction of T Cells and Cell Lines
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PBMCS were isolated by Ficoll-Paque (GE Healthcare, cat no. Cytvia 17-1440-03)
from buffy coats obtained from Sanquin Blood Bank). αβT cells were expanded from PBMCs using CD3/CD28 dynabeads (Thermo Fisher scientific, cat no. 40203D) and (1.7 × 103 IU/ml of MACS GMP Recombinant Human interleukin (IL)-7 (Miltenyi Biotec, cat no.130-095-361 ), and 1.5 × 102 IU/ml MACS GMP Recombinant Human IL-15 (Milteny Biotec, cat no. 130-095-762 ). HL60, RPMI 8226, and SSC9 stably expressing GFP-luciferase was generated by a previously described retroviral transduction protocol (30 (link)). The plasmid containing the GFP and luciferase transgenes was kindly provided by Jeanette Leusen (UMC Utrecht, Utrecht, Netherlands). The following cell lines were obtained from ATCC between 2010 and 2018, HL60 (CCL-240), RPMI 8226 (CCL-155), SCC9 (CRL-1629) and Daudi (CCL-213). Freestyle 293-F cells (R790-07) were obtained from Invitrogen. ML-1, HL60, RPMI 8226 and Daudi were cultured in RPMI (Gibco, cat no. 12017599), 10% FCS (Bodinco), 1% Pen/Strep (Invitrogen, cat no. 11548876). Freestyle 293-F in Freestyle expression medium (Gibco, cat no. 10319322). SCC9 in DMEM (Gibco, cat no. 31966047) 10% FCS, 1% Pen/Strep.
from buffy coats obtained from Sanquin Blood Bank). αβT cells were expanded from PBMCs using CD3/CD28 dynabeads (Thermo Fisher scientific, cat no. 40203D) and (1.7 × 103 IU/ml of MACS GMP Recombinant Human interleukin (IL)-7 (Miltenyi Biotec, cat no.
4
Characterization of AML Cell Lines
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The AML cell lines U937, MOLM-14, MV4-11, HL60, HEL, THP-1, NOMO-1, OCI-AML2, OCI-AML3 and NB4 were provided by collaborators or were purchased from ATCC or DSMZ (Braunschweig, Germany). All cell lines were cultured in RPMI-1640 medium containing 10% FBS (GIBCO, Life Technologies, Carlsbad, CA, USA), 2 μM l−1 glutamine, 100 U mL−1 penicillin, and 100 μg ml−1 streptomycin (GIBCO, Life Technologies, Carlsbad, CA, USA). The 293T cell line was purchased from ATCC and cultured in DMEM containing 10% FBS (GIBCO, Life Technologies, Carlsbad, CA, USA), 2 μM l−1 glutamine, 100 U mL−1 penicillin, and 100 μg mL−1 streptomycin (GIBCO, Life Technologies, Carlsbad, CA, USA). Daunorubicin (DNR) and cytarabine (ARA-C) were purchased from Selleck Chemicals LLC (Houston, TX, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively; SIRT6 chemical inhibitor [2,4-dioxo-N-(4-(pyridin-3-yloxyphenyl)-1,2,3,4-tetrahydroquinazoline-6-sulfonamide, henceforth named compound 1] was obtained from MolPort (Riga, Latvia).
5
Cell Line Culture and Treatment Protocols
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KG-1 (ATCC), HL-60 (ATCC), and MV-4-11 (ATCC) human cell lines were grown in IMDM medium (Thermo Fisher Scientific); THP-1 (ATCC) and SET-2 (ATCC) human cells were grown in RPMI (Thermo Fisher Scientific); and C3-luc cell line (22 (link)) was grown in DMEM (Thermo Fisher Scientific). Media were supplemented with 10% FBS (Atlanta Biologicals) and 1% penicillin-streptomycin (Thermo Fisher Scientific). Stable FOXM1-knockdown cell lines using the ATCC obtained parental cells were generated as described in ref. 23 (link). All cells were maintained at 37°C in 5% CO2. Cytarabine (MilliporeSigma) and ixazomib (Takeda) were dissolved in DMSO for cell culture experiments; Azacitidine (Wockhardt) and doxycycline (LKT Laboratories) were dissolved in PBS. By company recommendation, ixazomib was dissolved in 5% cyclodextran (MilliporeSigma) for the in vivo animal experiments.
6
Glycolysis Modulation in Cell Lines
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HL-60, K562, Jurkat, and NB-4 cells (ATCC, USA) were cultured in RPMI1640 (GIBCO, USA), 10% fetal bovine serum, 2 mM l -glutamine, 50 U/ml penicillin, and 50 μg/ml streptomycin. All these cell lines were authenticated and tested for mycoplasma contamination. Cells were treated with 400 μM cobalt chloride (CoCl2) (Amresco, USA) for 48 h to induce glycolysis, or with 1 mg/ml 2-deoxy-d -glucose (2-DG) (Sigma-Aldrich, USA) for 48 h to suppress glycolysis.
7
Megakaryocyte Differentiation from Cord Blood
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Cord blood was obtained from the Clinic of Gynecology of the Medical University of Vienna. All donors gave their written informed consent. CD34+ hematopoietic stem cells were isolated from cord blood with CD34 MACS magnetic beads (Miltenyi, Bergisch Gladbach, Germany). CD34+ cells were cultured with 50 ng mL−1 thrombopoietin, 1 ng mL−1 stem cell factor and interleukin‐3 (Miltenyi) in Stem Pro 34 medium (Invitrogen, Carlsbad, CA, USA) at 37 °C in 5% CO2 for 12 days to obtain mature culture differentiated megakaryocytes. Mature megakaryocytes were stained with CD41, CD14 and CD45 antibodies (eBioscience, San Diego, CA, USA), and their purity was analyzed by fluorescence‐activated cell sorting (FACS). Cultures containing > 3% contaminating leukocytes were used for our experiments. Cell lines CHRF, Meg‐01, HL‐60 and HepG2 were purchased from ATCC. They were grown in RPMI medium or Dulbecco's modified Eagles's medium (Invitrogen) supplemented with 10% fetal bovine serum and gentamicin (Gibco, Carlsbad, CA, USA) at 37 °C in 5% CO2.
8
Culturing Primary Alveolar and Airway Cells
9
Establishment of Leukemia Cell Lines
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MOLM13 were purchased from DSMZ (Braunschweig, Germany). MV4;11 and HL60 were purchased from ATCC (Manassas, VA, USA). OCI-AML3 were kindly provided by Mark Minden (Ontario Cancer Institute, Toronto, ON, Canada). MOLM13 luciferase (luc)/green fluorescent protein (gfp) cells were generated using lentiviral transduction. The lentiviral plasmid was generated by cloning the firefly luciferase sequence excised from pGL4.51 (Promega, Madison, WI, USA) into the pCDH-CMV-MCS-EF1-copGFP lentiviral vector (System Biosciences Inc., Mountain View, CA, USA).
MOLM13 p53 short hairpin ribonucleic acid (shRNA) cells were previously described.22 (link) BM-MSC were acquired in accordance with regulations and protocols approved by the Investigational Review Board of the University of Texas MD Anderson Cancer Center (MDACC). Informed consent was obtained in accordance with the Declaration of Helsinki.
MOLM13 p53 short hairpin ribonucleic acid (shRNA) cells were previously described.22 (link) BM-MSC were acquired in accordance with regulations and protocols approved by the Investigational Review Board of the University of Texas MD Anderson Cancer Center (MDACC). Informed consent was obtained in accordance with the Declaration of Helsinki.
10
Culturing HUVEC and HL-60 Cells
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Human umbilical vein endothelial cells (HUVEC) were obtained from Promo Cell (Heidelberg, Germany). Human monocytic leukemia cell line (HL-60) was purchased from ATCC (Manassas, VA, USA). HUVECs and HL-60 were cultured at a density of 5 × 105 cells/mL in Endothelial Cell Basal Medium2 (Promo cell, Heidelberg, Germany) and RPMI 1640 medium (Thermo scientific, Rockford, IL, USA). 10% heat-inactivated FBS and 100 U/mL penicillin G are supplemented in these medium. Cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2 and 95% air.
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