Effect of isolated neutrophils from Cd177 knockout mice post-SCI on microglia in a co-culture system

To investigate the transcriptional effects of CD177+ neutrophils on microglia in spinal cord injuries (SCI), we adapted a co-culture system containing isolated neutrophils from Cd177 WT or KO mice post-SCI and isolated microglia. Each group had a biological repeat (n=3). Briefly, we isolated neutrophils in injured lesions at 3 dpi from Cd177 KO or WT mice by magnetic-activated cell sorting (MACS) and co-cultured them with microglia. After co-culturing for 24h, We isolated microglia by fluorescence-activated cell sorting (FACS, gated on Ly6G-) for further testing.

为探究CD177阳性中性粒细胞对脊髓损伤（spinal cord injury, SCI）中小胶质细胞的转录调控效应，本研究构建了适配实验需求的共培养体系，体系包含脊髓损伤后Cd177野生型（wild type, WT）或敲除型（knockout, KO）小鼠分离得到的中性粒细胞，以及纯化分离的小胶质细胞。每组设置3次生物学重复（n=3）。简要而言，本研究于损伤后第3天（3 dpi），通过磁激活细胞分选（magnetic-activated cell sorting, MACS）从Cd177 KO及WT小鼠的损伤病灶中分离中性粒细胞，并将其与小胶质细胞进行共培养。共培养24小时后，通过荧光激活细胞分选（fluorescence-activated cell sorting, FACS，以Ly6G设门）分离小胶质细胞，用于后续实验检测。