The study was conducted using three HNSCC cell lines: H103 (ECACC, no. 06092001), FaDu (ATCC®, HTB-43™), and HPV positive—KB (ATCC®, CCL-17™). FaDu and KB cell lines were cultured in DMEM (Biowest, Nuaillé, France) supplemented with 10% fetal bovine serum (Biowest, France) and 1% of penicillin and streptomycin mixture (Merck, Darmstadt, Germany); H103 cells were cultured in 1:1 DMEM and Ham’s F12 (Biowest, France) nutrient medium with 10% fetal bovine serum, 1% of L-glutamine (Biowest, France) and 1% of penicillin and streptomycin mixture. All cells were cultured at 37 °C with 100% humidity at the 5% CO2 atmosphere.
Ccl 17
Manufactured by American Type Culture Collection
Sourced in United States
CCL-17 is a cell line derived from a human T-cell lymphoma. It is used for research purposes in the study of T-cell biology and related diseases.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Htb 43, by American Type Culture Collection (1 mentions)
Ham s f12, by Biowest (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
L glutamine, by Biowest (1 mentions)
Dulbecco modified eagle medium (dmem), by Biowest (1 mentions)
Pcs 201 018, by American Type Culture Collection (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Sas jcrb0260, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Scc25 crl 1628, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Crl 2974, by American Type Culture Collection (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Ach chloride, by Merck Group (1 mentions)
Htb 37, by American Type Culture Collection (1 mentions)
Antibiotic antimycotic solution, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
8 protocols using ccl 17
1
Head and Neck Cancer Cell Lines
2
In vitro Stimulation of Gingival Fibroblasts and Oral Cancer Cells
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Both human Gingival Fibroblast, GF (ATCC® PCS-201-018™) and Mouth Epidermal Carcinoma, KB (ATCC®, CCL-17) cell lines were grown as monolayers culture under standard conditions in DMEM medium containing 10% FCS. For in vitro stimulation, P. gingivalis (ATCC® 33277) and S. sanguinis (ATCC® 10556) bacterial cultures were allowed to grow in Luria-Bertani (LB) medium at 37 °C, and the optical density (OD) was measured at 578 nm (OD578). Bacteria were then harvested and washed twice in PBS, diluted to working solutions, and heat-inactivated at 68 °C for 20 min. For indicated experiments, bacteria were further centrifuged at 3000× g for 20 min. To separate the membrane components of the bacterial pathogens. Then, the heat-killed bacterial compounds were used to stimulate GF and KB cells corresponding to approximately 2 × 104 CFU/mL for S. sanguinis and P. gingivalis as well, per well. Whereas LPS concentrations of 100 ng/mL (Sigma-Aldrich; St. Louis, MO, USA) and LTA concentrations of 50 ng/mL were used for the stimulation of GF and KB cells.
3
Establishment of Human Oral Cancer Cell Lines
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The human oral squamous cell carcinoma (OSCC) cell line SAS (JCRB0260) was purchased from the Japanese Collection of Research Bioresources Cell Bank. SCC25 (CRL-1628) was obtained from the American Type Culture Collection (ATCC). OEC-M1 cell line was derived from oral cavity epidermal carcinoma [12 (link)], which is a generous gift from Prof. Jenn-Han Chen (National Defense Medical Center, Taiwan). KB drug-resistant cancer cells were purchased from the ATCC (CCL-17; Rockville, MA, USA). KB-7D cells were generated through etoposide (VP-16)-driven selection, which demonstrated topoisomerase II downregulation and multidrug resistance-associated protein overexpression. KB-tax cells were generated through taxol-driven selection. These drug-resistant cancer cells were kindly provided by Dr. Jang-Yang Chang (Cancer Research Division of National Health Research Institutes, Taiwan) [13 (link)]. Human oral cancer SAS, SCC25, and OEC-M1 cells were cultured in Roswell Park Memorial Institute 1640 medium. The culture medium was supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, and 2 mmol/L L-glutamine. The cells were grown at 37 °C in a humidified 5% CO2 incubator.
4
Optimized HCMV Virus Production Protocol
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MRC5 primary fetal lung fibroblasts (ATCC CCL-17) and ARPE-19 retinal pigment epithelial cells (ATCC CRL-2302) were purchased directly from ATCC, were grown in Gibco DMEM (MRC5) or Gibco DMEM/F-12 (ARPE-19) full media, supplemented with 10% (v/v) Fetal Bovine Serum (Cell Sera), 10 U/ml penicillin and 10 U/ml streptomycin and cultured in 5% CO2 at 37°C. All cells were regularly tested for mycoplasma contamination using PCR, and found to be negative. For vesicle-related experiments, confounding bovine vesicles were depleted from serum by centrifugation at 100,000xg for 1 hr at 4°C.
HCMV was reconstituted by electroporating MRC5 or ARPE-19 cells with bacterial artificial chromosomes (BAC) containing the AD169 genome kindly provided by Prof. Thomas Shenk (Yu et al., 2002 (link)), or Merlin kindly provided by Dr Richard Stanton (Stanton et al., 2010 (link)). Cellular supernatant was harvested and a sorbitol cushion (20% (v/v) D-sortbitol, 50 mM Tris pH 7.4) underlaid, prior to ultracentrifugation at 50,000xg for 1 hr at 4°C (Britt, 2010 (link)). Virus pellets were resuspended in full media, titred by TCID50 assay, and stored at −80°C. For HCMV infections, virus was added to cells at the specified multiplicity of infection (MOI) with gentle agitation over 3 hr, then aspirated and replaced with full media until harvesting.
HCMV was reconstituted by electroporating MRC5 or ARPE-19 cells with bacterial artificial chromosomes (BAC) containing the AD169 genome kindly provided by Prof. Thomas Shenk (Yu et al., 2002 (link)), or Merlin kindly provided by Dr Richard Stanton (Stanton et al., 2010 (link)). Cellular supernatant was harvested and a sorbitol cushion (20% (v/v) D-sortbitol, 50 mM Tris pH 7.4) underlaid, prior to ultracentrifugation at 50,000xg for 1 hr at 4°C (Britt, 2010 (link)). Virus pellets were resuspended in full media, titred by TCID50 assay, and stored at −80°C. For HCMV infections, virus was added to cells at the specified multiplicity of infection (MOI) with gentle agitation over 3 hr, then aspirated and replaced with full media until harvesting.
5
Culturing Folate Receptor-Positive Cancer Cells
6
Pharmacological Evaluation of Cell Lines
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All solvents were of analytical quality and obtained from VWR International. ACh chloride (purity > 99 %) was purchased from Sigma-Aldrich. Atropine (purity > 99 %) was obtained from TCI Chemicals. Galanthamine (purity > 99 %) was kindly provided by Janssen-Cilag. Cell lines used: HepG2 (ATCC HB-8065, clone H20) was obtained from Prof. Mersch-Sundermann, Freiburg, Germany, HaCaT (ATCC CCL-17) was a friendly gift from Prof. Fusenig, Heidelberg, Germany to A. H., and CaCo-2 cells (ATCC HTB-37, ACC 169) were kindly provided by Prof. Langer, Münster, Germany. Primary human normal dermal fibroblasts were resected from human foreskins, which were kindly provided by the University Hospital of Münster, Germany.
7
Propagation of Human Epidermal Carcinoma Cells
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KB (human epidermal carcinoma) cell line was purchased from the American Type Culture Collection (ATCC; CCL-17™). Cells were cultured in Eagle’s Minimum Essential Medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) and 1% Antibiotic Antimycotic Solution (Sigma-Aldrich, Saint Louis, MO, USA). For tumor induction, the cells were used after 6–8 passages and 85% confluence. The viability of the cells was always higher than 90%, as assessed by the trypan blue exclusion test.
8
Culturing and Maintaining Cancer Cell Lines
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The human lung LU-1 cancer (ATCC HTB-57 TM), lung A549 cancer, epidermoid carcinoma (KB, ATCC number CCL-17), hepatocellular carcinoma (HepG2, ATCC number HB-8065), breast (MCF7, HTB-22™) cancer cell lines, and human embryonic kidney (HEK-293, ATCC number CRL-1573) cell line were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). These cell lines were stored in liquid nitrogen, cultured in DMEM (Dulbeccos Modified Eagle Medium) medium with 7–10% fetal bovine serum (Gibco, USA), and 100 U per mL penicillin, and 100 μg mL−1 streptomycin (Gibco, USA) in a humidified atmosphere with 5% CO2 at 37 °C. The cells in the log phase are preferentially used for the tests.
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