All human T-ALL cell lines (KOPT-K1, DND41, JURKAT, SUP-T13, HPB-ALL, MOLT-4, MOLT-16, RPMI-8402, TALL-1, and LOUCY) were obtained from ATCC (Manassas, VA, USA) or DSMZ (Braunschweig, Germany). The creation of Ba/F3 cells transformed by TYK2-E957D was described previously.6 (link) JURKAT and KOPT-K1 cells overexpressing BCL2 were generated with the MSCV-IRES-GFP retroviral expression system. JURKAT and KOPT-K1 cells overexpressing BCLXL or MCL1 cDNA were generated with the pHAGE-CMV-IRES-ZsGreen lentiviral expression system. For additional information, see Supplementary Materials and Methods . These cells were maintained in RPMI-1640 medium (GIBCO, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin (Invitrogen, Waltham, MA, USA).
Kopt k1
Manufactured by American Type Culture Collection
Sourced in United States
The KOPT-K1 is a laboratory equipment designed for cell culture applications. It provides a controlled environment for the growth and maintenance of cells. The core function of the KOPT-K1 is to maintain optimal temperature, humidity, and atmospheric conditions necessary for cell cultivation.
Automatically generated - may contain errors
Lab products found in correlation
Jurkat, by American Type Culture Collection (9 mentions)
Molt 4, by American Type Culture Collection (8 mentions)
Ccrf cem, by American Type Culture Collection (6 mentions)
Hpb all, by American Type Culture Collection (5 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Tall 1, by American Type Culture Collection (4 mentions)
Loucy, by American Type Culture Collection (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Dnd41, by American Type Culture Collection (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Sup t1, by American Type Culture Collection (3 mentions)
Sup t13, by American Type Culture Collection (2 mentions)
Molt 16, by American Type Culture Collection (2 mentions)
Rpmi 8402, by American Type Culture Collection (2 mentions)
Hct116, by American Type Culture Collection (2 mentions)
Dnd41, by Leibniz Institute DSMZ (2 mentions)
Molt 3, by American Type Culture Collection (2 mentions)
F2442, by Merck Group (2 mentions)
Na934, by Merck Group (2 mentions)
12 protocols using kopt k1
1
Establishment and Maintenance of Human T-ALL Cell Lines
2
Establishment and Characterization of Human Cancer Cell Lines
3
Establishment and Maintenance of Human T-ALL Cell Lines
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All human T-ALL cell lines (KOPT-K1, DND41, JURKAT, SUP-T13, HPB-ALL, MOLT-4, MOLT-16, RPMI-8402, TALL-1, and LOUCY) were obtained from ATCC (Manassas, VA, USA) or DSMZ (Braunschweig, Germany). The creation of Ba/F3 cells transformed by TYK2-E957D was described previously.6 (link) JURKAT and KOPT-K1 cells overexpressing BCL2 were generated with the MSCV-IRES-GFP retroviral expression system. JURKAT and KOPT-K1 cells overexpressing BCLXL or MCL1 cDNA were generated with the pHAGE-CMV-IRES-ZsGreen lentiviral expression system. For additional information, see Supplementary Materials and Methods . These cells were maintained in RPMI-1640 medium (GIBCO, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin (Invitrogen, Waltham, MA, USA).
4
T-ALL Cell Line and PDX Generation
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The human leukemia cell lines CCRF-CEM, HPB-ALL, KOPT-K1, LOUCY, MOLT4, and SUP-T1 were purchased from the ATCC (Manassas, VA, USA) or Deutsche Sammlung von Mikroorganismen und Zellkulturen (Braunschweig, Germany). The mouse T-ALL cell lines LPN228 and LPN49 were generated from conditional knockout mice deficient in PTEN [27 (link)]. The cells were maintained in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum with penicillin and streptomycin (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C with 5% CO2 in a humidified incubator. T-ALL PDX cells CUL76, 6506870 and D115 were previously established and kindly provided Dr. Adolfo Ferrando (Columbia University, New York, NY, USA) and Dr. Marina Konopleva’s Lab (UTMDACC, Houston, TX) respectively [28 (link), 29 (link)].
5
Establishment and Characterization of Human Cancer Cell Lines
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Human colon cancer cell line HCT116 and human T-ALL cell lines Molt4, Molt3, Jurkat and KOPT-K1 were obtained from ATCC. HPBALL and DND41 human T-ALL cell lines were obtained from DSMZ (The Leibniz Institute). The CUTLL1 NOTCH1-dependent T-cell lymphoblastic cell line has been previously described (ref. 24 (link)). SHMT1 and SHMT2 were knocked out in HCT116 lines using CRISPR/Cas9 nickase method as previously described (ref. 4 (link),18 (link)). Adherent cell lines were subcultured in 5% CO2 at 37 °C using DMEM (CellGro 10–017; Mediatech) supplemented with 10% FBS (F2442; Sigma-Aldrich); suspension cell lines were subcultured in 5% CO2 at 37 °C in RPMI-1640 (11875; Gibco) with 10% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin. For all experiments, media supplemented with 10% dialyzed FBS was used (F0392; Sigma-Aldrich). Cell lines were regularly tested for mycoplasma. Antibodies were used according to their manufacturers’ directions. Anti-SHMT1 (12612) and SHMT2 (12762) were obtained from Cell Signaling Technologies (1:1,000 dilution). Anti-β-ACTIN (A3854) was obtained from Sigma-Aldrich (1:50,000 dilution). Secondary antibody coupled to horseradish peroxidase (NA934) was obtained from Sigma-Aldrich. Signal was detected using enhanced chemiluminescence (34578, Thermo Scientific).
6
Establishing Human T-ALL and BMSC Cultures
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Five human T-ALL cell lines (Loucy, Jurkat, CCRF-CEM, KOPT-K1, and DND-41) were obtained from ATCC (Manassas, USA) and maintained in RPMI-1640 medium (Sigma, Saint-Louis, USA) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 mg/mL streptomycin, and 100 U/mL penicillin at 37°C with 5% CO 2 . Human primary peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers from our hospital as previously described [25] and cultured in RPMI-1640 medium (Sigma) supplemented with 10% FBS. Human BMSCs (Shanghai Institute of Biological Sciences, Shanghai, China) were cultured in α-MEM (Sigma) containing 1% antibiotics and 10% FBS at 37°C with 5% CO 2 . BMSCs subjected to osteogenic differentiation were cultured in mineralization medium containing β-glycerophosphate (10 mmol/L), dexamethasone (10 -8 mol/L), and vitamin C (50 mg/L) for 14 days. The medium was changed every three days.
7
Characterization of T-ALL Cell Lines
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The human T-ALL cell lines CCRF-CEM, KOPT-K1, MOLT4, JURKAT, LOUCY, SUPT1
and the 293T cell line were purchased from American Type Culture Collection
(Manassas, VA, USA) and recently identified by DNA fingerprint. Two human
postnatal normal thymocyte samples were provided by Dr. Andrew Weng (Terry Fox
Laboratory, Canada). The mouse T-ALL cell lines (LPN248, LPN236, LPN228) were
generated from mouse Pten knock-out T-ALL models and LPN211 was
generated from Ink4a/Arf knock-out mice.30 (link) The CCRF-CEM-FFluc cell line was
obtained from Dr. Malcolm K. Brenner and was described previously.34 (link) The cell lines were cultured
in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine
serum (FBS) and 10 mM L-glutamine. 293T cells were cultured in
Dulbecco’s Modified Eagle Media (DMEM) with 10% FBS. Cells were
kept at 37°C in 5% CO2 and tested without cytoplasm
contamination.
and the 293T cell line were purchased from American Type Culture Collection
(Manassas, VA, USA) and recently identified by DNA fingerprint. Two human
postnatal normal thymocyte samples were provided by Dr. Andrew Weng (Terry Fox
Laboratory, Canada). The mouse T-ALL cell lines (LPN248, LPN236, LPN228) were
generated from mouse Pten knock-out T-ALL models and LPN211 was
generated from Ink4a/Arf knock-out mice.30 (link) The CCRF-CEM-FFluc cell line was
obtained from Dr. Malcolm K. Brenner and was described previously.34 (link) The cell lines were cultured
in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine
serum (FBS) and 10 mM L-glutamine. 293T cells were cultured in
Dulbecco’s Modified Eagle Media (DMEM) with 10% FBS. Cells were
kept at 37°C in 5% CO2 and tested without cytoplasm
contamination.
8
Human T-ALL Cell Line Culturing
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In the present study, human T-ALL cell lines, including TALL-1, KOPTK1, Jurkat, CCRF-CEM and Molt16, were purchased from the American Type Culture Collection (ATCC). The cells mentioned above were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin, (NanJing SunShine Biotechnology Co., Ltd.) and 100 µg/ml streptomycin (Sunshine Biotechnology, Nanjing, China). The cells were maintained in a humidified atmosphere with 5% CO2 at 37˚C.
Clinical samples were obtained from Zibo Central Hospital (Zibo, China) from February 2018 to April 2019. Blood samples were obtained from 20 pediatric patients (age range, 3.6-14 years; 10 males, 10 females) who were diagnosed with T-ALL and 20 healthy donors (age range, 3-15.2 years; 10 males, 10 females). All fresh blood samples were immediately separated into several portions, snap-frozen in liquid nitrogen and stored at -80˚C prior to protein and RNA extraction. The present study was approved by the Ethical Review Committee of Zibo Central Hospital. All participants and their legal guardians agreed to the use of their samples in the present study and written informed consents were obtained from all of the legal guardians of all participants.
Clinical samples were obtained from Zibo Central Hospital (Zibo, China) from February 2018 to April 2019. Blood samples were obtained from 20 pediatric patients (age range, 3.6-14 years; 10 males, 10 females) who were diagnosed with T-ALL and 20 healthy donors (age range, 3-15.2 years; 10 males, 10 females). All fresh blood samples were immediately separated into several portions, snap-frozen in liquid nitrogen and stored at -80˚C prior to protein and RNA extraction. The present study was approved by the Ethical Review Committee of Zibo Central Hospital. All participants and their legal guardians agreed to the use of their samples in the present study and written informed consents were obtained from all of the legal guardians of all participants.
9
Culturing Human T-ALL Cell Lines
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Human T-ALL cell lines (HPB-ALL, TALL-1, KOPTK1, Jurkat, CCRF-CEM, and Molt 4) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Molt 4-luciferase (Molt 4-luc) cells with stable luciferase expression were obtained from PerkinElmer (Santa Clara, CA, USA). T-ALL cells were cultured in Roswell Park Memorial Institute-1640 (RPMI-1640; Gibco, BRL, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; Gibco). All cells were incubated in a humidified incubator with 5% CO2 at 37°C. Cells from passages 2–4 were used for experiments.
10
Cell Line Characterization Protocol
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The human umbilical vein cell line, EA.hy926, human embryonic kidney cell line, HEK293, and T‐acute lymphoblastic leukaemia cell line, KOPTK1, were obtained from American Type Culture Collection (ATCC) and cultured according to the manufacturer's protocols.
Genomic DNA was isolated from cells using the Guide Cell and Tissue Genomic DNA Extraction Kit (Tiangen, Beijing, China). The genotypes of cell lines were determined by Taqman genotyping assay.
Genomic DNA was isolated from cells using the Guide Cell and Tissue Genomic DNA Extraction Kit (Tiangen, Beijing, China). The genotypes of cell lines were determined by Taqman genotyping assay.
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