Peripheral
blood was obtained
from healthy donors and collected in plastic Monovette EDTA tubes,
and the isolation of the mononuclear cells (i.e., human peripheral
mononuclear cells “PBMC”) was accomplished using Histopaque
(Sigma-Aldrich, St. Louis, MO, USA) as already reported.46 (link) Subsequently, 3 mL of blood was cautiously layered
over 3 mL Histopaque and centrifuged at 400g for
30 min at room temperature. The PBMC-containing layer at the interface
between blood serum and Histopaque was transferred into a new tube
and washed with PBS three times. The isolated cells were suspended
in Panserin 413 medium (PAN-Biotech, Aidenbach, Germany) supplemented
with 2.5% phytohemagglutinin M (PHA-M, Life Technologies, Darmstadt,
Germany). Finally, cell viability was measured using the resazurin
method as described above.
blood was obtained
from healthy donors and collected in plastic Monovette EDTA tubes,
and the isolation of the mononuclear cells (i.e., human peripheral
mononuclear cells “PBMC”) was accomplished using Histopaque
(Sigma-Aldrich, St. Louis, MO, USA) as already reported.46 (link) Subsequently, 3 mL of blood was cautiously layered
over 3 mL Histopaque and centrifuged at 400g for
30 min at room temperature. The PBMC-containing layer at the interface
between blood serum and Histopaque was transferred into a new tube
and washed with PBS three times. The isolated cells were suspended
in Panserin 413 medium (PAN-Biotech, Aidenbach, Germany) supplemented
with 2.5% phytohemagglutinin M (PHA-M, Life Technologies, Darmstadt,
Germany). Finally, cell viability was measured using the resazurin
method as described above.
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