RPMI-1640 Medium

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	supplemented with 10% fetal bovine serum (#IVT3008403, In Vitro Technologies, Australia) and 2 mM L-glutamine (#25030-081, Gibco®, USA)

Protocol tips
supplemented with 10% fetal bovine serum (#IVT3008403, In Vitro Technologies, Australia) and 2 mM L-glutamine (#25030-081, Gibco®, USA)

Publication protocol

Human prostate non-malignant PNT1a and prostate cancer 22RV1, LNCaP (clone FCG) and DU145 cell lines were obtained from the European Collection of Cell Cultures via CellBank Australia (Children's Medical Research Institute, Westmead, NSW, Australia). PNT1a and 22RV1 cells were maintained in RPMI-1640 medium (#R0883, Sigma-Aldrich, USA), supplemented with 10% fetal bovine serum (#IVT3008403, In Vitro Technologies, Australia) and 2 mM L-glutamine (#25030-081, Gibco®, USA). LNCaP cells were maintained in RPMI-1640 medium, supplemented with 2 mM L-glutamine, 10 mM HEPES (#H0887, Sigma-Aldrich, USA) and 1 mM sodium pyruvate (#S8636, Sigma-Aldrich, USA). DU145 cells were cultured in MEM medium (#M5650, Sigma-Aldrich, USA), supplemented with 10% fetal bovine serum, 2 mM L-glutamine and 1 mM sodium pyruvate. The prostate cell lines were incubated at 37°C with 5% CO2 in a Sanyo MCO-17AI humidified incubator (Sanyo Electric Biomedical Co., Ltd.). Cells were passaged at approximately 80% confluence, for detachment TrypLE™ Express (#12604-021, Gibco®, USA) was used. The PNT1a cell line was chosen as a non-malignant control for comparison to three cancer cell lines, 22RV1, LNCaP and DU145. Prostate cancer lines 22RV1 and LNCaP closely mimic aspects of the more common clinical disease, as they are AR-responsive and express PSA [17]. On the other hand, DU145 represents a late-stage prostate cancer with moderate metastatic potential , and is neither hormone-sensitive nor expresses PSA.

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