All cells were maintained at 37°C in humidified Steri-Cult CO2 incubators (Thermo Scientific). C4–2B cells (gift from L. Chung, Cedars-Sinai Medical Center, Los Angeles, CA), DU145 cells (ATCC) and 22Rv1 cells (gift from S. Plymate University of Washington) were maintained in RPMI-1640 Media (Gibco, Life Technologies) with 10% fetal bovine serum (Atlanta Biologicals). VCaP cells (purchased directly from ATCC; CRL-2876) were maintained in DMEM (ATCC) with 10% fetal bovine serum (Atlanta Biologicals). NCI-H660 (purchased directly from ATCC; CRL-5813) cells were maintained in HITES media (ATCC) supplemented with 5% FBS. All cell lines used in this project were validated through STR analysis using ATCC reference genomes and were routinely tested for mycoplasma using the MycoFluor Mycoplasma Detection Kit (Invitrogen).
Hites media
Manufactured by American Type Culture Collection
HITES media is a culture medium used for the isolation and cultivation of Haemophilus influenzae. It provides the necessary nutrients and growth factors required for the growth of this microorganism.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions)
Steri cult co2 incubators, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Mycofluor mycoplasma detection kit, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (1 mentions)
Human tumor dissociation kit, by Miltenyi Biotec (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Celltitre glo 3d cell viability assay, by Promega (1 mentions)
3 protocols using hites media
1
Cell Culture Maintenance Protocols
2
Maintaining NCI-H660 Cells and Dissociating LuCaP Tumors
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NCI-H660 cells (obtained directly from ATCC, CRL-5813) were maintained in HITES media (ATCC) with 5% fetal bovine serum (Atlanta Biologicals) in humidified Steri-Cult CO2 incubators (Thermo Scientific). LuCaP patient-derived xenograft tumors (established in-house; [12 (link)]) were harvested and dissociated using the human Tumor Dissociation Kit (Miltenyi Biotec) according to manufacturer’s protocol.
3
Comparative Cell Viability Assay
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LuCaP PDX cells (2.0X104) and NCI-H660 cells (5.0x103) were seeded in 96-well plates in either phenol red-free RPMI 1640 media (Gibco) supplemented with 5% charcoal/dextran treated fetal bovine serum (Atlanta Biologicals) and 1X pen/strep (Gibco) or HITES media (ATCC). Cells were treated 4–6 hours after seeding with vehicle, AMG-337 (50 nM) or cabozantinib (2.5 μM) in three replicate wells. Viability was assessed at time 0 and 72 hours post treatment using the CellTitre Glo 3D-Cell Viability Assay (Promega) according to manufacturer’s protocols.
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