Healthy peripheral-blood mononuclear cells (PBMCs): Buffy coat samples from the blood-component separation process were diluted with phosphate-buffered saline (1:1). Gradient centrifugation using Lymphoprep™ (STEMCELL Technologies, Vancouver, BC, Canada) was employed for PBMCs’ isolation. The PBMCs were collected and counted under a light microscope with a hemocytometer.
Buffy coat: EDTA whole-blood specimens were centrifuged at 1600× g for 15 min; the buffy coat layer between packed red blood cells and plasma was collected and stored at −80 °C. The cryopreserved buffy coat samples from OS patients and healthy donors were lysed with lysis buffer RA1 (Macherey Nagel, Düren, Germany).
Primary cells: Primary cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) with 10% (v/v) fetal bovine serum and maintained in a humidified atmosphere of 37 °C with 5% CO2 [21 (link)].
Buffy coat: EDTA whole-blood specimens were centrifuged at 1600× g for 15 min; the buffy coat layer between packed red blood cells and plasma was collected and stored at −80 °C. The cryopreserved buffy coat samples from OS patients and healthy donors were lysed with lysis buffer RA1 (Macherey Nagel, Düren, Germany).
Primary cells: Primary cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) with 10% (v/v) fetal bovine serum and maintained in a humidified atmosphere of 37 °C with 5% CO2 [21 (link)].
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