GSM7986002: Mildly degeneration, replicate 2, scRNA-seq; Homo sapiens; RNA-Seq
The nucleus pulposus tissues were dissociated into single cell suspension with a Singleron PythoNTM Automated Tissue Dissociator and sCelLiveTM Tissue Dissociation Mix. Cell viability was determined by trypan blue staining and evaluated by microscopy. Samples with >80% cell viability were continued for subsequent experiments. Single-cell suspensions were loaded onto microfluidic chips to obtain a single cell per  microwell. ScRNA-seq libraries were constructed by GEXSCOPE Single-Cell Sequencing Kit and Singleron Matrix Automated single-cell processing system. Individual libraries were diluted to 4 ng/µL and pooled for sequencing on Illumina novaseq 6000 with 150 bp paired end reads.