PC-3 (CRL-1425), LNCaP (CRL-1740), and MDA PCa 2b (CRL-2422) cell lines were obtained from the American Type Tissue Collection (ATCC, Manassas, VA) at the time of this work and authenticated by morphology, karyotyping, and short tandem repeat profiling-based approaches. Cell lines were also validated by ATCC for absence of mycoplasma contamination. PC-3 cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and L-glutamine (Thermo Fisher Scientific, Waltham, MA). LNCaP cells were grown in Roswell Park Memorial Institute (RPMI) medium with 10% FBS and 1% L-glutamine (Thermo Fisher Scientific). MDA PCa 2b cells were maintained in BRFF-HPC1 medium (Athena ES, Baltimore, MD) supplemented with 20% FBS. RC77 T/E cell line11 (link) was maintained in Keratinocyte-SFM medium with epidermal growth factor 1–53 and bovine pituitary extract (Thermo Fisher Scientific) on plates coated with FNC coating mix (Athena ES). All cell lines were maintained at 37°C and 5% CO2.
Protocol detail
Cell Line Maintenance Protocols for Prostate Cancer Research
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Bovine
Cell
Cell lines
Eagle
Epidermal growth factor
Fetal bovine serum
Glutamine
Keratinocyte
Mycoplasma
Pc 3 cells
Penicillin
Short tandem repeat
Streptomycin
Corresponding Organization : George Washington University
Other organizations : University of Maryland Eastern Shore, University of Hawaii System, University of Michigan–Ann Arbor, National Institute of Dental and Craniofacial Research, Uniformed Services University of the Health Sciences
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Variable analysis
independent variables
- Cell lines: PC-3 (CRL-1425), LNCaP (CRL-1740), and MDA PCa 2b (CRL-2422)
- Cell culture media: Dulbecco's Modified Eagle Medium (DMEM), Roswell Park Memorial Institute (RPMI) medium, and BRFF-HPC1 medium
- Outcomes (not explicitly mentioned)
- Fetal bovine serum (FBS) concentration (10% for PC-3 and LNCaP, 20% for MDA PCa 2b)
- Penicillin-streptomycin and L-glutamine (for PC-3 cells)
- L-glutamine (for LNCaP cells)
- Cell line authentication (morphology, karyotyping, short tandem repeat profiling)
- Absence of mycoplasma contamination
- Temperature (37°C) and CO2 concentration (5%)
- RC77 T/E cell line maintained on FNC coating mix
- Not explicitly mentioned
- Not explicitly mentioned
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