All fibroblasts were cultured in DMEM containing 4.5g/L glucose, 1.5g/L NaHCO3, 10% Fetal Bovine Serum, 4mM L-glutamine (Corning, Corning, NY), and 1% penicillin/streptomycin (Invitrogen, Waltham, MA). The methodology of our 7-day 3D-cell derived ECM (also recognized as CDM) production has been published previously [30 ]. At the completion of 3D matrix production, cells were rinsed twice with PBS and switched to a serum/EV-free DMEM (1% pen/strep, 4 μM glutamine) and were sustained for 48 hours at 37°C, to generate conditioned media (CM). Upon completion of the conditioning period, CM was removed to be processed for further experiments, and the cells and CDMs were lysed for protein extraction (see methods below for depiction of metabolite extraction). All PDAC cell lines used in this study were maintained in 4:1 DMEM (1 g/L glucose, 110 mg/mL sodium pyruvate) M3 Base (INCELL, San Antonio, TX) supplemented with 5% FBS, and 1% penicillin-streptomycin, until needed for experimental purposes.
Penicillin streptomycin
Manufactured by INCELL
Penicillin-streptomycin is a commonly used antibiotic combination solution for cell culture applications. It is a sterile liquid that contains the antibiotics penicillin and streptomycin.
Automatically generated - may contain errors
3 protocols using penicillin streptomycin
1
3D-cell Derived ECM Production and Conditioned Media
2
3D-Cell Derived ECM Protocol
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All fibroblasts were cultured in DMEM containing 4.5 g/L glucose, 1.5 g/L NaHCO3, 10% FBS, 4 mmol/L l -glutamine (Corning), and 1% penicillin/streptomycin (Invitrogen). The methodology of our 7-day three-dimensional (3D)-cell derived ECM (also recognized as CDM) production has been published previously (30 ). At the completion of 3D matrix production, cells were rinsed twice with PBS and switched to a serum/EV-free DMEM (1% pen/strep, 4 μmol/L glutamine) and were sustained for 48 hours at 37°C, to generate conditioned media (CM). Upon completion of the conditioning period, CM was removed to be processed for further experiments, and the cells and CDMs were lysed for protein extraction (see methods below for depiction of metabolite extraction). All PDAC cell lines used in this study were maintained in 4:1 DMEM (1 g/L glucose, 110 mg/mL sodium pyruvate) M3 Base (INCELL) supplemented with 5% FBS, and 1% penicillin-streptomycin, until needed for experimental purposes.
3
3D Cell-derived ECM Production and Conditioning
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All fibroblasts were cultured in DMEM containing 4.5g/L glucose, 1.5g/L NaHCO 3 , 10% Fetal Bovine Serum, 4mM L-glutamine (Corning, Corning, NY), and 1% penicillin/streptomycin (Invitrogen, Waltham, MA). The methodology of our 7-day 3D-cell derived ECM (also recognized as CDM) production has been published previously [33] . At the completion of 3D matrix production, cells were rinsed twice with PBS and switched to a serum/EV-free DMEM (1%pen/strep, 4 µM glutamine) and were sustained for 48 hours at 37°C, to generate conditioned media (CM). Upon completion of the conditioning period, CM was removed to be processed for further experiments, and the cells and CDMs were lysed for protein extraction (see methods below for depiction of metabolite extraction). All PDAC cell lines used in this study were maintained in 4:1 DMEM (1g/mL glucose, 110 mg/mL sodium pyruvate) M3 Base (INCELL, San Antonio, TX) supplemented with 5% FBS, and 1% penicillin-streptomycin, until needed for experimental purposes.
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