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Pe ca pj49

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The PE/CA-PJ49 is a laboratory equipment product. It serves a core function as a cell line, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Fetal bovine serum (fbs), by Gemini Bio (5 mentions) Dmem f12, by Thermo Fisher Scientific (3 mentions) L glutamine, by Thermo Fisher Scientific (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Hydrocortisone, by Thermo Fisher Scientific (2 mentions) Detroit 562, by American Type Culture Collection (2 mentions) Bicr 18, by Merck Group (2 mentions) Iscove s dmem, by Corning (2 mentions) Ampf str identifiler pcr amplification kit, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions) Hydrocortisone, by Merck Group (2 mentions) Dulbecco modified eagle medium (dmem), by Corning (2 mentions) L glutamine, by Corning (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Cal 27 cells, by American Type Culture Collection (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) Fugene hd, by Promega (1 mentions) Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions) Keratinocyte sfm, by Thermo Fisher Scientific (1 mentions)

6 protocols using pe ca pj49

1

HNSCC Cell Line Cultivation and Transfection

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All HNSCC cell lines were genotypically verified as previously described. [4 ] Cal27 cells were obtained from ATCC (Manassas, VA). PE/CA-PJ34clone12 and PE/CA-PJ49 cells were obtained from Sigma-Aldrich (St. Louis, MO). 686LN cells were obtained from Georgia Chen at MD Anderson Cancer Center (Houston, TX). Cal27 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Mediatech, Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS) (Gemini Bio-Products, West Sacramento, CA). 686LN cells were cultured in DMEM/F12 (Life Technologies, Grand Island, NY) supplemented with 10% FBS. PE/CA-PJ34clone12 and PE/CA-PJ49 were cultured in Iscove's Modification of DMEM (Mediatech Inc., Manassas, VA) supplemented with 10% FBS and 2 mM L-glutamine (Life Technologies, Grand Island, NY). All cells were maintained in an incubator at 37°C and 5% CO2. JSI-124 (Calbiochem, Billerica, MA) was dissolved in DMSO. Transfection was performed with Lipofectamine 2000 (Life Technologies, Grand Island, NY) or FuGENE HD (Promega Corporation, Madison, WI) according to the manufacturer’s instructions with 4 μg DNA diluted in Opti-MEM (Life Technologies, Grand Island, NY).
Peyser N.D., Du Y., Li H., Lui V., Xiao X., Chan T.A, & Grandis J.R. (2015). Loss-of-Function PTPRD Mutations Lead to Increased STAT3 Activation and Sensitivity to STAT3 Inhibition in Head and Neck Cancer. PLoS ONE, 10(8), e0135750.
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2

Establishing HNSCC Cell Lines

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The HPV(+) HNSCC cell lines UM-SCC47, 93-VU-147T, UPCI-SCC90, UD-SCC2, and UM-SCC104 were kindly provided by Dr. Randall Kimple (University of Wisconsin-Madison, Madison, WI). The HPV(+) HNSCC cell line UPCI-SCC152 was purchased from ATCC. All HPV(+) cell lines were maintained in DMEM with 10% FBS (Gemini Bio-Products) and 1% penicillin and streptomycin (Life Technologies) supplemented with 1% nonessential amino acids (Gibco). The HPV(−) cell lines SCC9 and Detroit562 were purchased from ATCC. PE/CA-PJ49 and PE/CA-PJ34 (clone 12) were purchased from Sigma-Aldrich. CAL33 was kindly provided by Dr. Gerard Milano (University of Nice, Nice, France). HSC-2 was obtained from the Health Science Research Resources Bank (Osaka, Japan). UM-SCC4, UM-SCC1, and TU138 were kindly provided by Thomas E. Carey (University of Michigan, Ann Arbor, MI). Normal oral keratinocytes-spontaneously immortalized (NOKSI) were kindly provided by Dr. Silvio Gutkind (University of California, San Diego, San Diego, CA). All HPV(−) cell lines were maintained in DMEM with 10% FBS and 1% penicillin and streptomycin. NOKSI cells were maintained in Keratinocyte SFM and supplemented with defined keratinocyte-SFM growth supplement (Gibco). CAL33, HSC-2, and Detroit562 express mutant PIK3CA (H1047R). All cell lines were authenticated by short tandem repeat testing (Genetica DNA Laboratories).
Brand T.M., Hartmann S., Bhola N.E., Li H., Zeng Y., O’Keefe R.A., Ranall M.V., Bandyopadhyay S., Soucheray M., Krogan N.J., Kemp C., Duvvuri U., LaVallee T., Johnson D.E., Ozbun M.A., Bauman J.E, & Grandis J.R. (2018). Cross-talk Signaling between HER3 and HPV16 E6 and E7 Mediates Resistance to PI3K Inhibitors in Head and Neck Cancer. Cancer research, 78(9), 2383-2395.
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3

Cell Line Verification and Maintenance

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Cal27 and Detroit 562 cells were obtained from ATCC (Manassas, VA). 686LN cells were obtained from Georgia Chen at MD Anderson Cancer Center (Houston, TX). BICR 18 and PE/CA-PJ49 cells were obtained from Sigma-Aldrich (St. Louis, MO). UMSCC cell lines were obtained from Thomas E. Cary at the University of Michigan (Ann Arbor, MI). HSC-2 cells were obtained from Hideo Niwa at Nihon University (Tokyo, Japan). Cal27, Detroit 562, HSC-2, UMSCC 47, and UMSCC 22A were maintained in DMEM (Corning, Corning, NY) containing 10% fetal bovine serum (FBS; Gemini Bio-Products, West Sacramento, CA). UMSCC 1 were maintained in DMEM containing 10% FBS and 0.4 μg/mL hydrocortisone (Sigma-Aldrich, St. Louis, MO). BICR 18 were maintained DMEM containing 10% FBS, 0.4 μg/mL hydrocortisone, and 2 mM L-glutamine (Life Technologies, Carlsbad, CA). 686LN were maintained in DMEM/F12 (Life Technologies, Carlsbad, CA) containing 10% FBS. PE/CA-PJ49 were maintained in Iscove’s DMEM (Corning, Corning, NY) containing 10% FBS and 2 mM L-glutamine. All cells were genotypically verified using the AmpFSTR Identifiler PCR Amplification Kit according to the manufacturer’s instructions (Life Technologies, Carlsbad, CA) and maintained at 37°C and 5% CO2.
Peyser N.D., Freilino M., Wang L., Zeng Y., Li H., Johnson D.E, & Grandis J.R. (2015). Frequent promoter hypermethylation of PTPRT increases STAT3 activation and sensitivity to STAT3 inhibition in head and neck cancer. Oncogene, 35(9), 1163-1169.
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4

Generation and Characterization of Cisplatin-Resistant Cell Lines

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All cell lines were cultured in DMEM (Fisher), 10% fetal bovine serum (Gemini Bio products) and 1% penicillin-streptomycin (Life Technologies). PE/CA-PJ49 and FaDu were purchased from Sigma-Aldrich and American Type Culture Collection (ATCC; Manassas, VA), respectively. All cell lines were authenticated periodically (at least every 6 months) via STR profiling (at the UC Berkeley Sequencing Core). Mycoplasma testing was performed periodically on all cell lines used in this study. Cetuximab resistant clones of PE/CA-PJ49 and FaDu were generated previously.4 (link) PE/CA-PJ49 and CAL33 cell lines were treated with 500nM cisplatin and surviving clones were selected and continually cultured in increasing doses up to 5uM. CAL33 cells were previously derived from a tongue SCC and were kindly provided by Dr. Gerard Milano (Centre Antoine-Lacassagne, Nice, France). Cisplatin resistance was confirmed by performing viability assays in comparison to their respective parental lines. JQ1 was provided by Dr. James Bradner (Dana Farber Institute) for in vitro studies and MZ1 was purchased from Tocris Bioscience (Bristol, UK).
Bhola N.E., Njatcha C., Hu L., Lee E.D., Shiah J.V., Kim M.O., Johnson D.E, & Grandis J.R. (2021). PD-L1 is upregulated via BRD2 in head and neck squamous cell carcinoma models of acquired cetuximab resistance. Head & neck, 43(11), 3364-3373.
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5

Cell Line Verification and Maintenance

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Cal27 and Detroit 562 cells were obtained from ATCC (Manassas, VA). 686LN cells were obtained from Georgia Chen at MD Anderson Cancer Center (Houston, TX). BICR 18 and PE/CA-PJ49 cells were obtained from Sigma-Aldrich (St. Louis, MO). UMSCC cell lines were obtained from Thomas E. Cary at the University of Michigan (Ann Arbor, MI). HSC-2 cells were obtained from Hideo Niwa at Nihon University (Tokyo, Japan). Cal27, Detroit 562, HSC-2, UMSCC 47, and UMSCC 22A were maintained in DMEM (Corning, Corning, NY) containing 10% fetal bovine serum (FBS; Gemini Bio-Products, West Sacramento, CA). UMSCC 1 were maintained in DMEM containing 10% FBS and 0.4 μg/mL hydrocortisone (Sigma-Aldrich, St. Louis, MO). BICR 18 were maintained DMEM containing 10% FBS, 0.4 μg/mL hydrocortisone, and 2 mM L-glutamine (Life Technologies, Carlsbad, CA). 686LN were maintained in DMEM/F12 (Life Technologies, Carlsbad, CA) containing 10% FBS. PE/CA-PJ49 were maintained in Iscove’s DMEM (Corning, Corning, NY) containing 10% FBS and 2 mM L-glutamine. All cells were genotypically verified using the AmpFSTR Identifiler PCR Amplification Kit according to the manufacturer’s instructions (Life Technologies, Carlsbad, CA) and maintained at 37°C and 5% CO2.
Peyser N.D., Freilino M., Wang L., Zeng Y., Li H., Johnson D.E, & Grandis J.R. (2015). Frequent promoter hypermethylation of PTPRT increases STAT3 activation and sensitivity to STAT3 inhibition in head and neck cancer. Oncogene, 35(9), 1163-1169.
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6

Culturing PE/CA-PJ49 Cell Line

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Cell lines were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Corning 10-013-CM) containing 10% fetal bovine serum (FBS; Gemini Bio-Products #900–108) and penicillin-streptomycin (Gibco 15140–122). PE/CA-PJ49 cells were purchased from Sigma-Aldrich. Cell lines were authenticated by short tandem repeat (STR) analysis performed by the University of California, Berkeley DNA Sequencing Facility at least once every 6 months.
O’Keefe R.A., Bhola N.E., Lee D.S., Johnson D.E, & Grandis J.R. (2020). Interleukin 6 is increased in preclinical HNSCC models of acquired cetuximab resistance, but is not required for maintenance of resistance. PLoS ONE, 15(1), e0227261.
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