HAC Media Information: Difference between revisions
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<xfeeds pubget="Y">doi:10.1210/jc.2008-0903</xfeeds> | <xfeeds pubget="Y">doi:10.1210/jc.2008-0903</xfeeds> | ||
<xfeeds pubget="Y">doi:10.1210/en.2008-1512</xfeeds> | |||
'''HAC15''' human adrenal carcinoma cells were plated in DMEM/F12 with L-glutamine and HEPES (Invitrogen) media with 10% cosmic calf serum (Fisher Scientific, Pittsburgh, PA), 1 µg/ml gentamicin (Invitrogen), and 1% insulin/transferrin/selenium premix (BD Bioscience) at 400,000 cells/well in 24-well tissue culture plates. Cells were allowed to adhere to plates and grow for 1 d, and media were replaced with a low serum media (0.1% cosmic calf serum). | |||
'''H295R''' human adrenal carcinoma cells were plated in DMEM/F12 (Invitrogen, San Diego, CA) media with 2.5% {nu} serum, and 1% insulin/transferrin/selenium premix (BD Biosciences, San Jose, CA) at 500,000 cells/well in 24-well tissue culture plates. Cells were allowed to adhere to plates and grow confluent for 1–2 d and then treated with vehicle dimethyl sulfoxide, angiotensin II (Sigma-Aldrich, St. Louis, MO), or test compound. At 24 and 48 h time points, 500 µl media were removed from each well and frozen at –20 C until needed for aldosterone and cortisol assays. Cells were lysed in buffer RLT (QIAGEN, Valencia, CA). Lysate was frozen at –80 C until RNA extraction. | |||
Revision as of 15:00, 24 January 2010
<xfeeds pubget="Y">doi:10.1210/jc.2008-0903</xfeeds>
<xfeeds pubget="Y">doi:10.1210/en.2008-1512</xfeeds>
HAC15 human adrenal carcinoma cells were plated in DMEM/F12 with L-glutamine and HEPES (Invitrogen) media with 10% cosmic calf serum (Fisher Scientific, Pittsburgh, PA), 1 µg/ml gentamicin (Invitrogen), and 1% insulin/transferrin/selenium premix (BD Bioscience) at 400,000 cells/well in 24-well tissue culture plates. Cells were allowed to adhere to plates and grow for 1 d, and media were replaced with a low serum media (0.1% cosmic calf serum).
H295R human adrenal carcinoma cells were plated in DMEM/F12 (Invitrogen, San Diego, CA) media with 2.5% {nu} serum, and 1% insulin/transferrin/selenium premix (BD Biosciences, San Jose, CA) at 500,000 cells/well in 24-well tissue culture plates. Cells were allowed to adhere to plates and grow confluent for 1–2 d and then treated with vehicle dimethyl sulfoxide, angiotensin II (Sigma-Aldrich, St. Louis, MO), or test compound. At 24 and 48 h time points, 500 µl media were removed from each well and frozen at –20 C until needed for aldosterone and cortisol assays. Cells were lysed in buffer RLT (QIAGEN, Valencia, CA). Lysate was frozen at –80 C until RNA extraction.
