We used HPC patient-derived xenograft cell lines (HPCM17 (link),8 (link) and HPCM27 (link),8 (link) cells), which were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium (Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS), 100 unit/mL penicillin, and 100 μg/mL streptomycin. MCC148c cells13 (link), established by patient-derived xenografts of cancer tissue from a lung squamous cell carcinoma patient5 (link), were maintained in DMEM supplemented with 10% FBS, 0.4 mg/mL hydrocortisone, 2.5 mM Y-27632 (Focus Biomolecules, Plymouth, PA, USA), and penicillin/streptomycin. DMEM supplemented with 10% FBS and penicillin/streptomycin was used to maintain 293 T cells (RIKEN BioResource Center, Kyoto, Japan). Het-1A cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and maintained in airway epithelial cell basal medium from the airway epithelial cell growth medium supplement pack (PromoCell, Heidelberg, Germany) supplemented with 4% FBS and penicillin/streptomycin. IMR-32 cells were purchased from ATCC and maintained in DMEM supplemented with 10% FBS and 0.1 mM non-essential amino acids (Thermo Fisher Scientific, Waltham, MA, USA). HSC-3 cells were purchased from RIKEN (Saitama, Japan) and maintained in Eagle’s minimal essential medium supplemented with 10% FBS and penicillin/streptomycin.
Imr 32 cells
Manufactured by American Type Culture Collection
Sourced in United States
The IMR-32 cell line is a human-derived neuroblastoma cell line. It is routinely used in various research applications, including the study of neurological disorders, cellular signaling pathways, and drug development.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by GeneDireX (3 mentions)
Sh sy5y cell, by American Type Culture Collection (2 mentions)
Minimum essential medium (mem), by GeneDireX (2 mentions)
Airway epithelial cell growth medium supplement pack, by PromoCell (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Y 27632, by Focus Biomolecules (1 mentions)
Rpmi 1640 medium, by Fujifilm (1 mentions)
Het 1a cell, by American Type Culture Collection (1 mentions)
293t cells, by RIKEN BioResource Center (1 mentions)
Penicillin streptomycin, by American Type Culture Collection (1 mentions)
Sk n be 2 cells, by American Type Culture Collection (1 mentions)
U118mg cells, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Kelly cells, by Merck Group (1 mentions)
Sk n as cells, by American Type Culture Collection (1 mentions)
Ccl 81, by American Type Culture Collection (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Sk n be 2c cells, by American Type Culture Collection (1 mentions)
11 protocols using imr 32 cells
1
Cell Culture Conditions for Patient-Derived Xenograft Models
2
Culturing Neuroblastoma and HEK293T Cells
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SK-N-BE (2) cells (ATCC) were cultured in Minimum Essential Medium supplemented with 100 nM L-glutamine, 1% penicillin-streptomycin, 1% 1 : 1 nonessential amino acid solution: Ham's F-12, and 15% fetal bovine serum (FBS). IMR-32 cells (ATCC) were cultured in Minimum Essential Medium supplemented with 100 nM L-glutamine, 1% penicillin-streptomycin, 1% nonessential amino acid solution, and 10% FBS. SH-SY5Y cells (ATCC) were cultured in Dulbecco's modified Eagle's medium (DMEM)/Nutrient Mixture F-12 Ham's supplemented with 100 nM L-glutamine, 1% penicillin-streptomycin, and 10% FBS. HEK293T cells were cultured in high-glucose DMEM supplemented with 100 nM L-glutamine, 1% penicillin-streptomycin, and 10% FBS (all from Sigma). All cells were cultured at 37°C in a humidified atmosphere with 5% CO2.
3
Culturing IMR-32 and U118-MG Cell Lines
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IMR-32 cells (ATCC, Manassas, VA) were cultured in EMEM (Gibco, Grand Island, NY) supplemented with 10% FBS (Gibco) and U118-MG cells (ATCC) were cultured in DMEM (Gibco) with 10% FBS. Both cell lines were incubated under standard growth conditions of 37º C and 5% carbon dioxide.
4
Neuroblastoma Cell Lines: Genetic Diversity
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Four neuroblastoma cell lines with different genetic makeup were used. Human neuroblastoma SK-N-AS cells (American Type Culture Collection (ATCC, Manassas, VA, USA)) were maintained in Dulbecco's modified Eagle's medium (DMEM, HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, 100 μg ml−1 streptomycin, and 0.1 mM non-essential amino acids. Human neuroblastoma IMR-32 cells (ATCC) were maintained in modified Eagle's medium (MEM, HyClone) supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, and 100 μg ml−1 streptomycin. Human neuroblastoma SH-SY5Y cells (ATCC) were maintained in 1 : 1 mixture of MEM and F-12 media supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, and 100 μg ml−1 streptomycin. Human neuroblastoma KELLY cells (Sigma-Aldrich, St Louis, MO, USA) were maintained in RPMI-1640 (HyClone) supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, 100 μg ml−1 streptomycin, and 2 mM glutamine. All cells were maintained in a 5% CO2 atmosphere at 37 °C and trypsin-passaged at 80% confluence.
5
Culturing Human Glioblastoma and Neuroblastoma Cells
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Human glioblastoma (GBM) cells (DBTRG-05MG, GBM cells) (Bioresource Collection and Research Center, BCRC-60380, Hsinchu, Taiwan) were maintained in RPMI-1640 medium (Gibco, #31800022, Waltham, MA, USA). Vero cells (American Type Culture Collection (ATCC), CCL-81, Manassas, VA, USA) and human neuroblastoma cells (IMR-32 cells, ATCC, CCL-127, Manassas, VA, USA) were cultured in MEM/EBSS medium (HyClone, #SH30008.02, Logan, UT, USA). Cell culture mediums were supplemented with 10% fetal bovine serum (FBS) (Gibco, A4766801, Waltham, MA, USA) and penicillin-streptomycin (P/S) (Gibco, #15140122, Waltham, MA, USA). PeV-A3 was from a virology group, Department of Microbiology, Kaohsiung Veterans General Hospital (27 (link)). PeV-A3 was propagated in Vero cells. The full genome sequence of PeV-A3 (strain VGHKKS-2007 accession #KM986843) has been characterized (28 (link)).
6
Lentiviral Knockdown of CSB in Neuronal Cell Lines
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SK-N-BE (2c) cells (American Type Culture Collection (ATCC) CRL-2271) were grown in a MEM/DMEM F12 medium supplemented with 10% Fetal Bovine Serum and 2 mM l -Glutamine.
IMR-32 cells (American Type Culture Collection (ATCC) CCL-127) were grown in a MEM medium supplemented with 10% Fetal Bovine Serum, 2 mM L-Glutamine, and Non-Essential Amino Acids.
ReNCell VM cells (SCC008, Sigma-Aldrich: St. Louis, MO, USA) were grown as an adherent monolayer on poly-ornithine (0.002%) laminin (2 mg/mL) coated tissue culture flasks in the presence of 20 ng/mL of human recombinant EGF and bFGF2 in DMEM:F12 medium with nutrients optimized for neural progenitor cell growth.
Cells in exponential growth phase were transduced with lentiviral shRNA particle (1 × 105 infectious units of virus—Santa Cruz Biotechnology, Santa Cruz, CA, USA), expressing sh-RNA targeting CSB or sh-RNA non-targeting control. Puromicin selection (2 μg/mL) is performed to achieve stable gene silencing.
Cells were treated with 5 μg/mL of Cytochalasin B for 24 h.
IMR-32 cells (American Type Culture Collection (ATCC) CCL-127) were grown in a MEM medium supplemented with 10% Fetal Bovine Serum, 2 mM L-Glutamine, and Non-Essential Amino Acids.
ReNCell VM cells (SCC008, Sigma-Aldrich: St. Louis, MO, USA) were grown as an adherent monolayer on poly-ornithine (0.002%) laminin (2 mg/mL) coated tissue culture flasks in the presence of 20 ng/mL of human recombinant EGF and bFGF2 in DMEM:F12 medium with nutrients optimized for neural progenitor cell growth.
Cells in exponential growth phase were transduced with lentiviral shRNA particle (1 × 105 infectious units of virus—Santa Cruz Biotechnology, Santa Cruz, CA, USA), expressing sh-RNA targeting CSB or sh-RNA non-targeting control. Puromicin selection (2 μg/mL) is performed to achieve stable gene silencing.
Cells were treated with 5 μg/mL of Cytochalasin B for 24 h.
7
Culture and Extraction of Cell Lines
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Two cell lines were used in our studies. One was the RAW264.7 mouse macrophage cell lines, purchased from Culture Collection and Research Center, Food Industry and Development Institute, Hsinchu, Taiwan. Cells were grown in Dulbecco's Modified Eagle Medium (Sigma) supplemented with 10% fetal bovine serum (Gibco Laboratories, Grand Island, NY) in a humidified atmosphere containing 5% CO2 at 37 °C. The second cell line was human neuroblastoma IMR-32 cells, obtained from the American Type Culture Collection (ATCC), were cultured in minimum essential medium (MEM) (Genedire X, USA) supplemented with 10% fetal bovine serum and 1% sodium pyruvate at 37•C in a controlled of 5% CO2 atmosphere. After 2-to 3-day growth, up to 70-80% full of 10 cm culture dish, the IMR-32 cultures were subcultured or collected and extracted out the nuclear proteins for EMSA use.
8
Cell Culture Protocols for Macrophage and Neuroblastoma
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Two cell lines were used in the present study. One was the RAW 264.7 mouse macrophage cell line purchased from Culture Collection and Research Center, Food Industry and Development Institute, Hsinchu, Taiwan. The cells were grown in Dulbecco's modi ed Eagle medium (Sigma) supplemented with 10% fetal bovine serum (Gibco Laboratories, Grand Island, NY) in humidi ed atmosphere containing 5% CO 2 at 37 °C. The second cell line was human neuroblastoma IMR-32 cells, obtained from the American Type Culture Collection (ATCC), which were cultured in minimum essential medium (MEM) (Genedire X, USA) supplemented with 10% fetal bovine serum and 1% sodium pyruvate at 37 ℃ in a controlled atmosphere of 5% CO 2 . After 2-to 3-day growth that lled up to 70-80% of a 10-cm culture dish, the IMR-32 cultures were subcultured or collected, and nuclear proteins were extracted for performing the electrophoretic mobility shift assay (EMSA). Chemicals LPS (Escherichia coli O111: B4) and DEX were purchased from Calbiochem® (Detroit, MI) and Sigma Chemical Co. (St. Louis, MO), respectively.
9
Modulation of STAT1/STAT3 Signaling in IMR-32 Cells
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IMR-32 cells were obtained from the American Type Culture Collection (Rockville, MA). Cell culture media components were obtained from Invitrogen Life Technologies (Carlbad, CA). Antibodies for STAT1, STAT3, β-actin, α-tubulin, β-tubulin and heterogeneous nuclear ribonucleoprotein A1 (hnRNP), and the oligonucleotides containing the consensus sequence for STAT1 and STAT3 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). The antibody for 4-hydroxynonenal was purchased from Abcam (Cambridge, MA). Antibodies for phosphotyrosine-701 STAT1 (pY701-STAT1), phosphoserine-727 STAT1 (pS727-STAT1), phosphotyrosine-705 STAT3 (pY705-STAT3), and phosphoserine-727 STAT3 (pS727-STAT3) were purchased from Cell Signaling Technologies (Danvers, MA). The reagents for EMSA assay were obtained from Promega (Madison, WI). Polyvinylidene difluoride (PVDF) membranes were obtained from Bio-Rad (Hercules, CA, USA). Chroma Spin-10 columns were obtained from Clontech (Mountain View, CA). Vinblastine (Vb), colchicine (Col), cytochalasin D (Cyt D), α-lipoic acid (LA) and all other reagents were of the highest quality available and were purchased from Sigma (St. Louis, MO).
10
Culturing Mouse Macrophages and Human Neuroblastoma Cells
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Two cell lines were used in the present study. One was the RAW 264.7 mouse macrophage cell line purchased from Culture Collection and Research Center, Food Industry and Development Institute, Hsinchu, Taiwan. The cells were grown in Dulbecco’s modified Eagle medium (Sigma) supplemented with 10% fetal bovine serum (Gibco Laboratories, Grand Island, NY) in humidified atmosphere containing 5% CO2 at 37 °C. The second cell line was human neuroblastoma IMR-32 cells, obtained from the American Type Culture Collection (ATCC), which were cultured in minimum essential medium (MEM) (Genedire X, USA) supplemented with 10% fetal bovine serum and 1% sodium pyruvate at 37 °C in a controlled atmosphere of 5% CO2. After 2- to 3-day growth that filled up to 70–80% of a 10-cm culture dish, the IMR-32 cultures were subcultured or collected, and nuclear proteins were extracted for performing the electrophoretic mobility shift assay (EMSA).
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