Pa tu 8902

The human carcinoma cell lines HCC1143 (breast carcinoma), PA-TU-8902 (pancreas adenocarcinoma), DU-145 (prostate carcinoma), and NCI-H460 (lung carcinoma) were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures (Germany). MV3 cells (human metastatic melanoma) were kindly provided by Anka Dahl, University Hospital, Hamburg-Eppendorf (Germany). All cell lines except PA-TU-8902 were cultivated under standard cell culture conditions (37°C, 95% relative humidity, 5% CO₂) in RPMI-1640 medium (Sigma-Aldrich, Buchs, Switzerland) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Sigma-Aldrich, Switzerland), 2 mM L-Glutamine (Sigma-Aldrich, Switzerland), and Penicillin-Streptomycin (Sigma-Aldrich, Switzerland; Penicillin 10,000 units/mL and 10 mg/mL Streptomycin). Dulbecco's Modified Eagle's Medium (DMEM, Sigma-Aldrich, Switzerland) supplemented with the ingredients mentioned above was used to cultivate PA-TU-8902 cells. The cells were maintained in adherent, exponential growth in 25 cm² culture flasks (TPP, Faust Laborbedarf AG, Switzerland) and cells from subconfluent monolayer were harvested by brief exposure to trypsin-EDTA solution (Sigma-Aldrich, Switzerland).

Human breast carcinoma cell lines MCF-7 and HCC1143, pancreas adenocarcinoma cell line PA-TU-8902, prostate >carcinoma cell line DU145 and lung carcinoma cell line NCI-H460 were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany).
In general RPMI 1640 medium, supplemented with 2 mM L-Glutamine and 1% Penicillin-Streptomycin and FCS (either 10% or 1% according to the assay) was used for monocyte/macrophage, NCI-H460, HCC1143 and DU145 cell culture. For MCF-7 cells we used Eagle´s MEM, supplemented with 10% FCS, 2 mM L-Glutamine, 1% Penicillin/Streptomycin and 1 mM Sodium Pyruvate and for PA-TU-8902 cells Dulbecco´s MEM High Glucose supplemented with 1 mM Sodium Pyruvate, 2 mM L-Glutamine, 1% Penicillin/Streptomycin and either 10% or 1% FCS was used (all reagents from Sigma, Buchs, Switzerland).
Cell lines were kept under standard culture conditions (37°C, 5% CO₂ and 95% humidity), tumour cell conditioned media were obtained by 24 h incubation of 90% confluent tumour cells in 10 ml of fresh culture medium in 25 cm² cell flasks. Cell free supernatants were carefully harvested, filtered through 0.5 μm filters and stored in aliquots at −80°C.

Human breast carcinoma cell lines HCC1937 and HCC1143, pancreas adenocarcinoma cell line PA-TU-8902, prostate carcinoma cell line DU145 and lung carcinoma cell line NCI-H460 were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany).
HCC1937, HCC1143, DU145 and NCI-H460 cells were cultured in RPMI-1640 supplemented with 10% fetal calf serum, 2 mM L-glutamine, and 1% Penicillin – Streptomycin (Sigma-Aldrich). PA-TU-8902 cells were cultured in Dulbecco’s MEM High Glucose (Sigma-Aldrich) supplemented with 2 mM L-Glutamine, 1 mM Sodium Pyruvate, 10% fetal calf serum and 1% Penicillin – Streptomycin in a humidified atmosphere with 5% CO₂ at 37°C. Cell lines were maintained in exponential growth and cells from subconfluent monolayers were harvested by trypsin-EDTA (Sigma-Aldrich) to carry out the experiments. For measurement of the parameters, the cell cultures were used within 4–6 weeks after thawing.