SK-N-AS, SK-N-FI, SK-N-BE(2)-C, SH-EP, SH-SY5Y, IMR-32, H460, A549, DU145, HCT116, HT29, PC3, SKOV3, CHP-134, Kelly, MDA-MB-231, and HEK-293T cells were purchased from ATCC by the Children’s Cancer Institute Australia Tumour Bank, and maintained in Dulbecco’s modified Eagle’s medium (Invitrogen, Australia) with 10% fetal calf serum. The human lung fibroblast cells MRC-5 and WI-38 (ATCC) were cultured in Alpha-MEM media with 10% FCS. Non-MYCN-amplified human neuroblastoma cell lines SH-EP MYCN-3 and SHEP-tet21N, which are genetically modified to overexpress or repress human MYCN cDNA when exposed to doxycycline, were generously supplied by Professor Jason Shohet (Texas Children’s Cancer Center, Houston). The shMYCN SK-N-BE(2)-C cell line are genetically modified to repress human MYCN expression by MYCN shRNA when exposed to doxycycline, and SK-N-BE(2)-C cell line was used as the parental cell line. The shMYCN SK-N-BE(2)-C cell line was derived by GenScript, Piscataway NJ. All cell lines used were authenticated by CellBank Australia (Westmead, NSW), to be free from mycoplasma, and were cultured at 37 °C with 5% CO2 in a humidifier incubator.
Chp134
Manufactured by American Type Culture Collection
Sourced in United States
The CHP134 is a high-performance cell culture incubator designed for precise temperature, humidity, and gas control to support the growth and maintenance of a wide variety of cell lines. The core function of this equipment is to provide a stable and optimized environment for cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (8 mentions)
Streptomycin, by Thermo Fisher Scientific (8 mentions)
Sk n as, by American Type Culture Collection (6 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Imr 32, by American Type Culture Collection (5 mentions)
Sms san, by American Type Culture Collection (5 mentions)
Nbl s, by American Type Culture Collection (5 mentions)
Sk n be, by American Type Culture Collection (4 mentions)
Sh sy5y, by American Type Culture Collection (3 mentions)
Sk n sh, by American Type Culture Collection (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Nb 39 nu, by American Type Culture Collection (2 mentions)
Lan 5, by American Type Culture Collection (2 mentions)
Pf 477736, by Merck Group (2 mentions)
Sk n fi, by American Type Culture Collection (1 mentions)
Sk n be 2 c, by American Type Culture Collection (1 mentions)
Sh ep, by American Type Culture Collection (1 mentions)
Wi 38, by American Type Culture Collection (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
9 protocols using chp134
1
Cell Lines for Neuroblastoma Research
2
Neuroblastoma Cell Line Transfection
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Neuroblastoma cell lines CHP-126, CHP-134, SH-SY5Y, SK-N-SH, LAN-1, and IMR-32 were procured from ATCC, USA. NB cells were cultured in F12/DMEM medium (Solarbio, Beijing, China) with 10% FBS, at 37°C and 5% CO2 incubator. DKC1 si-RNA, miR-326-5p inhibitor, miR-326-5p mimic, DKC1 mimic, and corresponding control vector (Jiuruiwu, Henan, Zhengzhou, China) were transfected into neuroblastoma cells, respectively, with Lipofectamine 3000 reagent (Semmerfeld, USA), and the operation was carried out strictly in accordance with kit instructions.
3
Neuroblastoma Cell Lines and Inhibitors
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The human NB cell lines CHP134, NB-39-nu, SKN-BE, and SMS-SAN were obtained from the American Type Culture Collection (Manassas, VA, USA) and the RIKEN Bioresource Cell Bank, Tohoku University Cell Resource Center (Miyagi, Japan) and grown in RPMI1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 50 µg/mL penicillin, and 50 µg/mL streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). CHK1i (PF-477736), ATMi (Ku55933), and DNA-PKi (NU7441) were obtained from Sigma-Aldrich St. Louis, MO, USA. Stock solutions were made in dimethyl sulfoxide (DMSO).
4
Neuroblastoma Cell Line Culturing
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NB cell lines (SK-N-AS, SK-N-BE(2), SK-N-SH, SJ-NB-10, NBL-S, CHP-212, IMR-32, NGP, Kelly, LAN-5, LA1-55N, CHP-134) used in this study have been previously utilized by our laboratory [50 (link),51 (link),52 (link)] and were purchased from American Type Culture Collection (ATCC, www.atcc.org ) or were generously provided by Susan Cohn (The University of Chicago Children’s Hospital, Chicago, IL, USA) or John Maris (Children’s Hospital of Philadelphia, Philadelphia, PA, USA). NB cells were maintained in RPMI-1640 media (Corning Life Science, Corning, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Life Technologies, Grand Island, NY, USA), 1% L-glutamine, 1% sodium pyruvate, and 1% non-essential amino acids and antibiotic/antimycotic solution (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C in 5% CO2.
5
Neuroblastoma Cell Culture and Inhibitor Assays
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The human neuroblastoma cell lines CHP134, NBLS, SKN‐BE, SK‐N‐AS, IMR32, and SMS‐SAN were purchased from the American Type Culture Collection and the RIKEN Bioresource Cell Bank, Tohoku University Cell Resource Center.24 , 25 Cells were cultured in RPMI 1640 medium supplemented with 10% heat‐inactivated FBS, 50 µg/mL penicillin, and 50 µg/mL streptomycin (Thermo Fisher Scientific). PF‐477736 (a selective inhibitor of CHK1) and gemcitabine (GEM) or trametinib (GSK1120212, a selective inhibitor of MEK1/2) were purchased from Sigma Aldrich or Chemitek, respectively.
6
Characterization of Neuroblastoma Cell Lines
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Human NB cell lines (NGP, CHP134, SK-N-AS and SK-N-SH) were obtained from the American Type Culture Collection (Manassas, VA, USA) and RIKEN Bioresource Cell Bank, Tohoku University Cell Resource Center (Miyagi, Japan). The cells were cultured in RPMI 1640 (Wako, Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 100 μg/mL penicillin/streptomycin (Sigma-Aldrich, St. Louis, MO, USA). Cells were cultured at 37 °C in a 5% CO2 incubator. All cell lines that we used in this study were tested and authenticated via STR assay, compared to the database at https://web.expasy.org/cellosaurus . The absence of mycoplasma contamination was confirmed using a Mycoplasma PCR Detection set (Takara Bio, Kusatsu, Shiga, Japan). These analysis was performed within 6 months when this work was completed.
7
Neuroblastoma Cell Line Cultivation
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We obtained the human neuroblastoma cell lines SKN-BE, NBLS, IMR32, SMS-SAN, and CHP134 from the American Type Culture Collection (Manassas, VA, USA) and the RIKEN Bioresource Cell Bank, Tohoku University Cell Resource Center (Miyagi, Japan) [19 (link),32 (link),33 (link)]. All cell lines were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 50 µg/mL penicillin, and 50 µg/mL streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). We purchased NU7441, a selective inhibitor of DNA-PKcs, from Sigma Aldrich (St. Louis, MO, USA).
8
Neuroblastoma Cell Lines Validation
9
Culturing Human Neuroblastoma Cell Lines
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The human NB cell lines NB-39-nu, SMS-SAN, NBLS, CHP134, SH-SY5Y and SK-N-AS, and SK-N-BE were purchased from the American Type Culture Collection (Manassas, VA, USA) and the RIKEN Bioresource Cell Bank, Tohoku University Cell Resource Center (Miyagi, Japan). Cells were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 50 μg/ml penicillin, and 50 μg/ml streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). 293FT cell line (Thermo Fisher Scientific) was maintained in DMEM medium with 10% FBS, 50 μg/ml penicillin, 50 μg/ml streptomycin, and 500 μg/ml Geneticin (Thermo Fisher Scientific). Cells were incubated at 37°C in a 5% CO2 humidified atmosphere (MCO-175-PJ, PHC, Tokyo, Japan). PF-477736 (henceforth, CHK1i) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Stock solutions were made in dimethyl sulfoxide (DMSO).
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