ST004
Homo sapiens
Protocols: Ascites was collected from 30 patients with high-grade serous ovarian carcinoma undergoing primary surgery at the University Hospital in Marburg. Informed consent was obtained from all patients according to the protocols approved by the local ethical committee. Adherent cells from ascites were positively selected on primaria 6 well dishes (3x10^6 cells per well in 2 ml PBS (PAA)) followed by 6 washing steps with PBS. Finally the cells were cultured for 6d in humidified incubator at 37 °C, 5 % CO2 in 2ml autologous ascites. Cells were treated with control (DMSO), L165 1 uM final concentration or ST247 300 nM final concentration for 1d. Cells were washed with PBS, supernatant was removed completly, then 1 ml of peqGOLD TriFast (PEQLAB) was added to each well and transfered to a 1,5 ml Eppi. Following 5 min of incubation at RT 200 ul trichloromethane were added to the solution. The probes were mixed thoroughly every 3 minutes during a 10 minute incubation at RT. Probes were centrifuged at 13.000 g for 15 min at 4 degrees C then the aquouse top phase was transfered to a new eppi containing 600 ul of ice cold 2-propanol. Probes were mixed and centrifuged for 10 min at 13.000 g, 4 degrees C. The supernatant was removed and the pelleted RNA was washed twice with 75 % Ethanol for 5 minutes at 13.000 g, 4 degrees C. The pellet was dried and dissolved in 15 ul of RNase free water following quality control using Xcalibur (Thermo scientific). Libraries were created using the Illumina Truseq mRNA kit used according to the manufacturer's instructions.
ENA first public
2015-06-01
ENA last update
2016-11-08
External Id
SAMEA3175751
INSDC center alias
Institute for molecular and tumor biology Philipps University Marburg Germany
INSDC center name
Institute for molecular and tumor biology Philipps University Marburg Germany
INSDC first public
2015-06-01T16:20:34Z
INSDC last update
2016-11-08T11:55:44Z
INSDC status
public
Submitter Id
E-MTAB-3167:ST004
broker name
ArrayExpress
cell type
tumor associated macrophages
clinical history
ascites collected from 30 patients with high-grade serous ovarian carcinoma undergoing primary surgery at the University Hospital in Marburg
common name
human
individual
OC54bTAM
sample name
E-MTAB-3167:ST004