A GD2-positive murine NB cell line NXS2 [18 (link)] and CHO cell line [19 (link)] (ATCC, Wesel, Germany) were cultured in Dulbecco's modified Eagle's medium supplemented with stable glutamine, 4.5 g/l glucose (DMEM; PAN-Biotech, Aidenbach, Germany), 10% (v/v) fetal calf serum (FCS), 100 U/ml penicillin and 0.1 mg/ml streptomycin (1× P/S; PAA, Pasching, Austria). Hybridoma cells producing murine anti-Id Ab ganglidiomab [17 (link)] were cultured in serum-free DMEM with stable glutamine and 4.5 g/l glucose supplemented with 1× non-essential amino acids (PAA, Pasching, Austria) and 50 μM β-mercaptoethanol (Sigma Aldrich, Steinheim, Germany). The human NB cell line LA-N-1 [20 (link)] was cultured in RPMI (PAN-Biotech, Aidenbach, Germany) supplemented with 4.5 g/l glucose, 2 mM stable glutamine (PAA, Pasching, Austria), 10% (v/v) FCS and 1× P/S. The genetically engineered NK-92-scFv(ch14.18)-zeta cell line (NK-92tr), expressing a GD2-specific chimeric antigen receptor derived from ch14.18, was kindly provided by Prof. W. Wels (Georg-Speyer Haus, Frankfurt, Germany) and cultured as previously described [21 (link)].
Cho cell line
Manufactured by American Type Culture Collection
Sourced in Germany, United States
The CHO cell line is a commonly used mammalian cell line derived from Chinese Hamster Ovary (CHO) cells. It is a well-established system for the expression and production of recombinant proteins.
Automatically generated - may contain errors
Lab products found in correlation
β mercaptoethanol, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by PAN Biotech (1 mentions)
Streptomycin, by PAN Biotech (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Penicillin, by Harvard Bioscience (1 mentions)
Streptomycin, by Harvard Bioscience (1 mentions)
Non essential amino acids (neaa), by Harvard Bioscience (1 mentions)
Sodium pyruvate, by Harvard Bioscience (1 mentions)
N acetyl l alanyl l glutamine, by Harvard Bioscience (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Collagen type 1 solution, by Nitta Gelatin (1 mentions)
L glutamine, by Fujifilm (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Fujifilm (1 mentions)
Caco 2 cell, by American Type Culture Collection (1 mentions)
5 protocols using cho cell line
1
Culturing Murine and Human NB Cell Lines
2
FRET Biosensor Cell Lines for Tau
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The Tau RD P301S FRET Biosensor cell line (CRL-3275) was provided by Marc Diamond. HEK293T cells were ordered from ATCC (CRL-11268). Cells were maintained in DMEM supplemented with 10% FBS and penicillin–streptomycin at 37 °C and 5% CO2 in a humidified atmosphere. CHO cell line was obtained from ATCC (CCL-61) and cultured in DMEM/F12 supplemented with 10% FBS and penicillin–streptomycin.
3
PET Imaging of PD-L1 Expression in CHO Tumors
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The Chinese hamster ovary (CHO) cell line was obtained from American Type Culture Collection. A CHO cell line with constitutive PD-L1 expression (CHO-hPD-L1) was generated in our laboratory and described previously (23 (link)). Details of the cell culture, flow cytometry analysis for PD-L1 expression, and in vitro assays are provided in the supplemental materials.
For PET imaging, nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mice bearing CHO-hPD-L1 or CHO tumors were intravenously injected with approximately 7.4 MBq (∼0.17 μg) of 68Ga-NOTA-WL12 in 200 μL of saline, and PET images were acquired at 30, 60, and 120 min. To establish in vivo specificity, animals were coinjected with 50 μg of unlabeled WL12. Image analysis and PD-L1 immunohistochemistry of the CHO-hPD-L1 and CHO tumors are described in the supplemental materials.
For PET imaging, nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mice bearing CHO-hPD-L1 or CHO tumors were intravenously injected with approximately 7.4 MBq (∼0.17 μg) of 68Ga-NOTA-WL12 in 200 μL of saline, and PET images were acquired at 30, 60, and 120 min. To establish in vivo specificity, animals were coinjected with 50 μg of unlabeled WL12. Image analysis and PD-L1 immunohistochemistry of the CHO-hPD-L1 and CHO tumors are described in the supplemental materials.
4
Transduction of Prostate Cancer Cell Lines
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The prostate cancer cell line PC3, LNCaP as well as the CHO cell line were purchased from American Type Culture Collection and have not been further authenticated. By FACS analysis an expression of PSCA was not detected in PC3 or LNCaP. PSMA expression was detectable in LNCaP but also not in PC3 cells. Therefore, PC3 cells were transduced with the open reading frame (orf) encoding PSCA or PSMA or both. LNCaP cells were transduced with orf encoding PSCA. For in vivo analysis the PC3 cell line expressing PSCA and PSMA was further modified to express the gene encoding firefly luciferase (Tu-Luc). Transduction was performed using a lentiviral packaging system as described previously [e.g. 15]. All cell lines were cultured in RPMI 1640 medium completed with 10% FCS, 100 U/ml penicillin and 100 μg/ml streptomycin, 2 mM N-acetyl-L-alanyl-L-glutamine, 1% non-essential amino acids and 1 mM sodium pyruvate (Biochrom). Cells were maintained at 37 °C in a humidified atmosphere of 5 % CO2.
5
Cellular Uptake and Metabolism of Phytochemicals
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The following materials were purchased from the listed sources: CHO cell line and Caco-2 cells from the American Type Culture Collection (Rockville, MD, USA); Dulbecco’s modified Eagle’s medium (DMEM) containing 4 mM L-glutamine from Wako Chemicals (Osaka, Japan); penicillin–streptomycin (10,000 U/mL and 10 mg/mL in 0.9% sodium chloride, respectively) from Gibco (Gaithersburg, MD, USA); fetal calf serum (FCS) from Life Technologies (Grand Island, NY, USA); non-essential amino acids (NEAA) from Cosmobio (Tokyo, Japan); [3H]-glucose (specific radioactivity: 21.2 Ci/mmol), [3H]-fructose (5.0 Ci/mmol), [3H]-L-leucine (142 Ci/mmol) from GE Healthcare (Fairfield, CT, USA), [3H]-L-glutamic acid (46 Ci/mmol), and [3H]-glycyl-sarcosine ([3H]-Gly-Sar) (0.2 Ci/mmol) from Moravek (Brea, CA, USA); and a type-I collagen solution from Nitta Gelatin (Osaka, Japan). The phytochemicals used for this study are shown in Table A1 . All other chemicals used were of reagent grade.
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