All reagents, unless otherwise specified, were purchased from Sigma-Aldrich (St. Louis, MO, USA). 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino (polyethylene glycol)-2000] (DSPE-PEG-2000 Amine) and 1,2-distearoyl-sn-glycero-3-phosphocholin (DSPC), which were used to synthesize the particles, were purchased from Avanti Polar Lipids (Alabaster, AL, USA). MDA-MB-231 breast cancer cells, Hs578T breast cancer cells, and MIA-Paca-2 pancreatic cancer cells were purchased from ATCC (HTB-26, HTB-126, and CRL-1420) (Manassas, VA, USA). A LIVE⁄DEAD® Viability/Cytotoxicity Kit was used to analyze cell viability (Invitrogen, Carlsbad, CA, USA), and a Cell Counting Kit-8 (CCK-8) was obtained from Dojindo (Rockville, MD, USA). A dialysis bag (MW cut-off of 6–8 kDa) was purchased from Spectrumlabs (Piraeus, Greece) to investigate the drug release behavior of the metal particles.
Mia paca 2 pancreatic cancer cells
Manufactured by American Type Culture Collection
Sourced in United States
MIA PaCa-2 are pancreatic cancer cells derived from a primary tumor. They are used for in vitro research and testing purposes.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Mda mb 231 breast cancer cells, by American Type Culture Collection (1 mentions)
Live dead viability cytotoxicity kit, by Thermo Fisher Scientific (1 mentions)
Htb 126, by American Type Culture Collection (1 mentions)
Htb 26, by American Type Culture Collection (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
1 2 distearoyl sn glycero 3 phosphoethanolamine n amino polyethyleneglycol 2000 dspe peg 2000 amine, by Avanti Polar Lipids (1 mentions)
Buffered formalin phosphate, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
Glass bottom dishes, by MatTek (1 mentions)
Anti il 17ra, by Amgen (1 mentions)
Anti il17a, by Amgen (1 mentions)
Anti il17f, by Amgen (1 mentions)
Abt 888, by Enzo Life Sciences (1 mentions)
Mycoalert plus mycoplasma detection kit, by Lonza (1 mentions)
Hy 13030, by MedChemExpress (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
Bxpc 3, by American Type Culture Collection (1 mentions)
Cfpac 1, by Thermo Fisher Scientific (1 mentions)
5 protocols using mia paca 2 pancreatic cancer cells
1
Synthesis and Evaluation of Therapeutic Nanoparticles
2
Culturing Pancreatic and HeLa Cells
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MIA PaCa-2 pancreatic cancer cells are purchased from ATCC. The HeLa Kyoto EB3-EGFP cells are purchased from Biohippo. Cells are cultured in Dulbecco’s Modified Eagle Medium (DMEM, ATCC) with 10% fetal bovine serum (FBS, ATCC) and 1% penicillin/streptomycin (Thermofisher Scientific). The cells are seeded in glass-bottom dishes (MatTek Life Sciences) with 2 mL culture media and then incubated in a CO2 incubator at 37 °C and 5% CO2 concentration. Cells are grown to about 50% confluency and are directly used for live-cell imaging or fixed with 10% buffered formalin phosphate (Thermofisher Scientific) for imaging.
3
Targeting IL-17 Axis in Pancreatic Cancer
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The animal experiments complied with ethical regulations for animal research and was approved (reference number no.42502-2-1266 uniMD) by the Institutional Animal Care and Use Committee of the Magdeburg University Hospital. Animals were maintained, and experiments were conducted following institutional guidelines. Antibody (500 µg/mouse) i.p. anti-IL-17RA, anti-IL-17A, anti-IL-17F (1:1:1; kindly provided by Amgen, Germany), or PBS was applied to mice, according to the timetable (see Figure 2 ). We obtained MIA PaCa-2 pancreatic cancer cells from the American Type Culture Collection (ATCC, Manassas, VA). L3.6pl is a metastatic pancreatic cell line generated by our lab [8 (link)] and the Panc02 cell line is a murine pancreatic cancer cell line. Cells in culture were treated with recombinant human respective mouse IL-17A (20 ng/mL, Peprotech, Rocky Hill, CT, USA; Biolegend, San Diego, CA, USA) or PBS in the tests.
4
Authenticating and Validating Pancreatic Cancer Cell Lines
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Panc1, BxPC3, and MiaPaCa2 pancreatic cancer cells were purchased from the American Type Culture Collection (Manassa, VA, USA). Cells were cultured under recommended conditions. All cell lines were authenticated by short tandem repeat DNA profiling at the UAB Heflin Center for Genomic Science (Birmingham, AL, USA). All pancreatic cancer cell lines used were tested for mycoplasma using MycoAlertTM PLUS Mycoplasma Detection Kit (Lonza, Walkersville, MD, USA) and results were negative. Olaparib (HY-10162, MedChem Express, Monmouth Junction, NJ, USA), JQ1 (a generous gift from Dr. Bradner; HY-13030, MedChem Express), and veliparib (a generous gift from Dr. Yang; ABT-888, Enzo Life Sciences, Farmingdale, NY, USA) were dissolved in DMSO.
5
Pancreatic Cancer Cell Line Cultivation
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CFPAC-1, HPAF-II and MIAPaCa2 pancreatic cancer cells were obtained from the American Type Culture Collection. S2-013 is a cloned subline of a human pancreatic tumour cell line, SUIT-2 (derived from liver metastasis). S2-013 and T3M4 cell lines were provided by M. A. Hollingsworth (Eppley Institute, The University of Nebraska Medical Center). The pancreatic cancer cell lines S2-013, HPAF-II, CFPAC-1, T3M4, MIAPaCa2 and KPC1245, as well as the stellate cell lines CAF-0911, primary CAF-0906, CAF-1003 and CAF-1016, human pancreatic stellate cells (HPSs) and HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1:100 antibiotic–antimycotic (Gibco). The low pH of the media was adjusted in the range ~6.9–7.1 by adding 1 g l−1 NaHCO3, and the control pH was set by using 3.7 g l−1 NaHCO3. The cell lines were routinely tested for mycoplasma contamination and confirmed by short tandem repeat (STR) profiling at the University of Arizona Genetics Core (UAGC) every six months or sooner.
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