CSF samples were centrifuged at 500g at 4℃ for 10 minutes to precipitate the CSF cells. The supernatant was carefully aspirated and 2 mL of red blood cell (RBC) lysis buffer was added to the precipitated cells to remove the RBC from the puncture injury. At 15-25°C, the cells were centrifuged at 500g for 5 min. After the supernatant was abandoned, the cells were washed twice with phosphate-buffered saline (PBS), and finally, the cells were re-suspended with 1mL PBS. Take 10 μL of the re-suspension, and stain with Trypan blue, then detect the proportion of surviving cells under the microscope and calculate the cell concentration. The concentration of cell suspension was adjusted to 1×105/mL using PBS. Employing the Singleron Matrix® platform, single-cell suspensions were deposited onto a microwell chip. Barcoding beads were retrieved from the microwell chip to synthesize cDNA through mRNA reverse transcription. The cDNA libraries were prepared through PCR amplification, enzymatic fragmentation, and adapter ligation using the GEXSCOPE® Single Cell RNA Library Preparation System according to the manufacturer's protocols (Singleron GEXSCOPE Single Cell RNA-seq Library Kit, Singleron Biotechnologies). Individual libraries were sequenced on Illumina NovaSeq 6000 with 150 bp paired end reads.