THP-1 suspension cellsCell aggregationTransfection efficiencyPMA pre-maturationAdherent cells such as mouse RAW cells or human cancer U2OS cells are beneficial to DNA transfection, with 20%-60% transfection efficiency. However, this DNA transfection is rarely used on suspension cells due to its low ...
Aspirate PMA-containingCulture mediawithout disturbing the cells (cells should be adherent after the 16h incubation period). 5. Wash cells with 300μL/well of 1× PBS for 5min at room temperature. Repeat this procedure. 6. Add 200μL/well of freshCulture mediaand incubate plates/cells for...
We designed and synthesized three small interfering RNA sequences based on the DUSP1 mRNA sequence, transfected siRNA-DUSP1 into post-adherent THP-1 cells, and observed the effect by fluorescence microscopy after 24 h (Fig. 2D). We then detected the efficiency of knockdown of the three small...
Upon PMA treatment the suspension culture changes and cells become adherent to glass and plastics and show an increase in mitochondrial and lysosomal numbers and in cytoplasmic to nuclear ratio, as well as an altered differentiation dependent on cell surface markers in a pattern similar to monocyte-...
After incubation, nonattached cells were removed by aspiration, and the adherent cells were washed with RPMI 1640 three times. THP-1 cells in RPMI 1640 without PMA were used as control (undifferentiated) cells. Flow cytometry analysis Expression of CD14 and CD147 on the surface of the ...
Briefly, THP-1 cells (26105/well) were infected withM.tuberculosis(26105CFU/ml)for3h(T0),thesupernatants werecollectedandmonolayerswashedthoroughlywithphosphate buffersaline(PBS)toremoveextracellularadherentmycobacteria. For the T0 wells, THP-1 cells were lysed with 100ml of 0.5% Triton X-100 and ...
Upon PMA treatment the suspension culture changes and cells become adherent to glass and plastics and show an increase in mitochondrial and lysosomal numbers and in cytoplasmic to nuclear ratio, as well as an altered differentiation dependent on cell surface markers in a pattern similar to monocyte-...
To solve this problem, we should ensure the THP-1 cells density (the appropriate density is 2–8×105 cells/mL.) and supplement high quality serum during cell culture. Furthermore, we could transfer the suspension cells to a new well plate and discard the adherent cells, when the number ...
THP-1 cells as non-adherent were transferred to a collection tube and centrifuged under the same conditions as PC12 cells. After removing the supernatant from PC12 cell cultures, a fresh medium was added, and the cell suspension was pressed twice through a 0.7 mm needle to break cell clumps...
While DMEM is selected for adherent cells, RPMI is widely used for suspension cells [68,69,70]. In line with this, RPMI is the recommended culture medium by the American Type Culture Collection (ATCC) and Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) for THP-1 cells. ...