Isolation of PBMCs
PBMCs were isolated by using a modified method of GmeligMeyling
and Waldmann.
23 Briefly, venous blood (10 mL)
was obtained in heparin-containing tubes containing equal
amount of physiological saline (1:1) by aseptic vein puncture
from healthy human volunteers who were nonsmokers, had
fasted overnight, and did not take any medicine or supplements.
Written informed consent was provided for protection
of the rights and welfare of human subjects involved in this
research. The diluted blood was then centrifuged at 400× g
for 20 minutes after carefully layered on lymphoprep. The
monocytes, formed as a distinct band at the medium interface,
were collected carefully using a Pasteur pipette. The collected