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Cell isolation and culture

XC Xinxin Chen
YW Yanqiu Wang
YQ Yicheng Qi
JY Jiqi Yan
FH Fengjiao Huang
MZ Mengxi Zhou
WW Weiqing Wang
GN Guang Ning
YZ Yulin Zhou
SW Shu Wang
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Peripheral blood samples were collected from GD patients and healthy controls. The peripheral blood mononuclear cells (PBMCs) were isolated via Ficoll-Hypaque density gradient centrifugation (Sigma Aldrich, St. Louis, MO, USA). Monocytes were purified from Ficoll-enriched PBMCs using human CD14 microbeads (Stemcell Technologies, Canada). The purification of monocytes was carried out according to the manufacturer’s instructions. Freshly isolated PBMCs or CD14+ monocytes were cultured in RMPI 1640 supplemented with 10% fetal bovine serum, 1% penicillin at 37 °C and in a 5% CO2 atmosphere.

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