Human g csf

The MAC-CYP cell generation was slightly modified from a previous report [25 (link)]. Briefly, after lysis of the red blood cells, bone marrow nucleated cells were collected and used for purification of CD11b⁺Gr1⁺ monocytes using the MACS technique. Gr1⁺ cells were positively selected using APC-conjugated anti-Gr1 antibody and anti-APC Microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). The positively selected cells were cultured in RPMI medium 1640 (Invitrogen) containing 10% FBS, human GM-CSF (R&D Systems), human G-CSF (R&D Systems), and human IL-13 (R&D Systems), each at 100 ng/mL for seven days. After seven days’ culture, the cells were transduced with indicated lentiviral vectors at a multiplicity of infection (MOI) of 5. Twenty-four hours later, the virus was removed, and culture media were replenished. The cells were cultured for another 24 h and examined for transduction efficiency under a fluorescence microscope. When necessary, the above transduction procedure was repeated one more time before use.