We are isolating human PBMCs (Ficoll-Paque) from fresh human blood samples and after 24hours we wish to treat the cells with a range of chemotherapy concentrations and perform a WST-1 (cell viability assay). As these cells are suspension cells, does anyone have a recommendation as to how to add the chemotherapy? I have read that you can add the desired concentration directly into the well, but we generally make up the treatments in fresh culture media and first remove the old media, rinse the cells and then add treatment.
Hello Caria Fourie,
Unlike adherent cells, suspension cells may be treated within 6-8 hours after plating. You need not wait for 24 hours. You may add the chemotherapeutic drug directly into the wells containing the cell suspension in concentrated form such that the desired concentration is achieved.
Alternatively, if you have a microplate compatible centrifuge, you may centrifuge the plate at 2000rpm for 5 mins at 4°C, carefully pipette out the old media and add fresh media containing the chemotherapeutic drug. You need to be careful as there is a risk of losing cells by using this method.
Best.
- Badges
- Science method