GSM8508987: CT_scTCR; Mus musculus; OTHER
An appropriate amount of human cardiac septal tissue was taken and placed into the serum-free medium with sterile forceps, and the tissue was minced with sterile scissors and filtered through a 40 μm filter to obtain the cell suspension. Subsequently, an appropriate amount of type II collagenase was added, and digestion was carried out in a 37 ℃ water bath for 20 min to terminate the enzyme digestion, and the residual collagenase was washed away by centrifugation-discarding the supernatant and resuspending with culture medium. 300g-500g for 5 min-8 min to remove the cellular debris. Finally, erythrocyte lysis was performed using erythrocyte lysis solution. The single cell suspension was loaded onto a 10X Chromium A Chip and the scRNA-Seq libraries were constructed following the protocol of the 10X genomics Chromium Single Cell Immune Profiling Solution. After the RT reaction, droplets were broken and barcoded-cDNA was purified with DynaBeads, followed by PCR-amplification. Sequencing libraries of single-cell transcriptomes suitable for Illumina sequencing platform were constructed after partial cDNA fragments and splicing. The remaining cDNA was enriched for the immune receptor (TCR), and the enriched products were amplified by PCR to construct a sequencing library suitable for the Illumina sequencing platform.