Colorectal adenocarcinoma cell line COLO 205 (ATCC #CCL-222), prostate adenocarcinoma cell lines PC3 (ATCC #CRL-1435) and DU145 (ATCC #HTB-81), and breast adenocarcinoma cell line MDA-MB-231 (ATCC #HTB-26), were purchased from American Type Culture Collection (Manassas, VA, USA). COLO 205 and PC3 cells were cultured in RPMI 1640 cell culture medium (Invitrogen, Grand Island, NY, USA), DU145 cells were cultured in EMEM cell culture medium (Invitrogen), and MDA-MB-231 cells were cultured in DMEM cell culture medium (Invitrogen). Media was supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) PenStrep, all purchased from Invitrogen. HUVECs were purchased from Lonza (Basel, Switzerland) and used from passages 4–5. HUVECs were cultured in Medium 199 (Invitrogen) and supplemented with EGM endothelial cell growth medium SingleQuots kit (Lonza). COLO 205, PC3, DU145, MDA-MB-231, and HUVECs cells were incubated under humidified conditions at 37 °C and 5% CO2, and were not allowed to exceed 90% confluence.
Egm endothelial cell growth medium singlequots kit
Manufactured by Lonza
The EGM endothelial cell growth medium SingleQuots kit is a supplemental kit for cell culture media. It contains a set of essential growth factors and supplements required for the optimal growth and maintenance of endothelial cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Huvecs, by Lonza (2 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Mda mb 231, by Thermo Fisher Scientific (1 mentions)
Colo205, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Rpmi 1640 cell culture medium, by Thermo Fisher Scientific (1 mentions)
Medium 199, by Thermo Fisher Scientific (1 mentions)
Dmem cell culture medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Huvecs, by Thermo Fisher Scientific (1 mentions)
Du145, by Thermo Fisher Scientific (1 mentions)
Colo205, by American Type Culture Collection (1 mentions)
2 protocols using egm endothelial cell growth medium singlequots kit
1
Cell Culture Protocol for Cancer Cell Lines
2
Coculture of hPSC-ECs and HPTECs
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HUVECs (Lonza) were cultured in EBM (Lonza) supplemented with EGM Endothelial Cell Growth Medium SingleQuots Kit (Lonza) and used at passage 3-6. The coculture procedures for HUVEC-HPTEC were the same as HKMEC-HPTEC coculture described above using HUVEC media. Human pluripotent stem cells (hPSC) (WTC line, a gift from Dr. Murry, University of Washington) was first differentiated to posterior-like ECs according to the protocol adapted from Palpant et al. (41) and Redd et al. (44) with low Activin A and high BMP4 for 10 days. Then these cells were trypsinized and plated into co-culture device (at center well) to allow for attachment and growth into confluency. HPTECs were plated into the outer well of the device at day 11, and co-cultured with hPSC-EC for additional two days in hPSC-EC media (41) . At day 13, five devices were fixed with PFA for immunostaining, and additional 5 devices were lysed for RNA collection. count. For other culture conditions where HKMECs were confluent, the average cell size was determined by field of view/total DAPI count.
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