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Cell purification and culture

ZH Zikun Huang
RS Rigu Su
ZD Zhen Deng
JX Jianqing Xu
YP Yiping Peng
QL Qing Luo
JL Junming Li
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The peripheral blood samples (5 mL) were collected from 22 healthy donors. Individuals with cancer, allergic diseases, immune-compromised conditions, diabetes or other infectious diseases such as HBV, HCV and HIV infection were excluded. After sample collection, PBMCs were isolated by density gradient centrifugation. To develop MDMs, the monocytes were purified using anti-CD14 magnetic beads (Miltenyi Biotec, USA) and the purified monocytes were cultured in RPMI 1640 medium contains 10% Human serum and 0.05% Glutamine (Sigma, USA) for 7 days at 5% CO2 and 37 °C14. The MDMs were identified by morphologic observation and flow cytometric analysis followed by anti-CD68 staining.

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