Selected article for: "fetal calf serum and flow cytometry"
Author: Ranjbar, Shahin; Haridas, Viraga; Jasenosky, Luke D.; Falvo, James V.; Goldfeld, Anne E.
Title: A Role for IFITM Proteins in Restriction of Mycobacterium tuberculosis Infection
  • Document date: 2015_10_22
    • ID: zy82bo7q_31
    Snippet: PBMCs were isolated by Ficoll-Hypaque (Pharmacia) density gradient centrifugation. Monocytes were enriched by positive selection with CD14 microbeads (STEMCELL Technologies) and then cultured at 1 × 10 6 cells per well in six-well plates in RPMI-1640 medium with 2 mM L-glutamine (BioWhittaker), supplemented with 5% heat-inactivated human AB serum (Atlanta Biologicals) and 50 ng/ml recombinant human GM-CSF (R&D Systems). After 5 days, supernatant.....
    Document: PBMCs were isolated by Ficoll-Hypaque (Pharmacia) density gradient centrifugation. Monocytes were enriched by positive selection with CD14 microbeads (STEMCELL Technologies) and then cultured at 1 × 10 6 cells per well in six-well plates in RPMI-1640 medium with 2 mM L-glutamine (BioWhittaker), supplemented with 5% heat-inactivated human AB serum (Atlanta Biologicals) and 50 ng/ml recombinant human GM-CSF (R&D Systems). After 5 days, supernatants were replaced with GM-CSF-free medium prior to experimental analysis. More than 98% of the adherent cells obtained with this technique were CD14 + macrophages, as verified by flow cytometry. THP-1 cells were obtained from ATCC and cultured in RPMI-1640 medium supplemented with 10% fetal calf serum (FCS, BioWhittaker). The 293T and A549 cells were obtained from ATCC and were maintained in DMEM (BioWhittaker) supplemented with 10% FCS.

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