Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham

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	supplemented with 10% FBS and antibiotics (all from Sigma, St. Louis, MO)

Protocol tips
supplemented with 10% FBS and antibiotics (all from Sigma, St. Louis, MO)

Publication protocol

Human prostate carcinoma DU145 cells were cultivated in DMEM-F12 HAM medium supplemented with 10% FBS and antibiotics (all from Sigma, St. Louis, MO). In order to establish the invasive DU145 cell sub-sets, mi-croporous membranes in a Boyden chamber were used (Corning; pore diameter – 8 mm; membrane diameter – 6.5 mm). Native DU145 cells were seeded at the density of 300 cells per mm2 onto the membrane and allowed to transmigrate for 48 hours. Afterwards, the cham-bers were placed into another well and the progeny of DU145 cells that precipitated onto the original well bot-tom was propagated (giving rise to DU145_48 subset). The “second wave” of transmigrating DU145 cells was collected in the next well for 24 hours and their progeny was propagated (DU145_72 subset). Before further anal-yses, the DU145 sub-sets were cultivated for at least 15 generation times (5 passages at 1:8)

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham below.

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