作者
Chen Li,Zhourui Wu,Liqiang Zhou,Jingliang Shao,Xiao Hu,Wei Xu,Yilong Ren,Xingfei Zhu,Weihong Ge,Kunshan Zhang,Jiping Liu,Runzhi Huang,Jing Yu,Dandan Luo,Xuejiao Yang,Wenming Zhu,Rongrong Zhu,Changhong Zheng,Yi Eve Sun,Liming Cheng
摘要
For single cell RNA sequencing dataset, the 10× Genomics platform was used to perform massive single cell mRNA profiling from mouse spinal cord samples collected at 6 different time points after Spinal Cord Injury (crush injury), i.e., 4h, 1d, 3d, 7d, 14d, and 38d post injury. Uninjured spinal cord samples are included as well. Both male and female C57BL/6 mice were used in the analyses, 10mm-long spinal cord segments encompassing the lesion site were dissociated into single cells through our proprietary method developed based on published studies1,2. For non-injury controls, male and female spinal cord samples were sequenced separately, and male and female scRNA-seq data overlapped rather well. For bulk RNA sequencing, 1 cm long spinal cord segments centered on leison core, average of 20 mg, at 15min, 1d, 3d, 7d, 14d, 28d, 42d after injury, were homogenized in 1 ml TRIzol (Invitrogen), and mRNA was extracted and purified by RNeasy Mini Kit (QIAGEN) according to the manufacturer’s instructions. The qualities of RNA were determined by Agilent 2100. 1-3 μg qualified RNA were subjected to Illumina V2 RNAseq library construction, followed by Hiseq2000 SE50bp Sequencing.