GSM5548128: bone marrow cells, T1D; Mus musculus; RNA-Seq
The femur and tibia were dissected. After removing the muscle and periosteum on the outer surfaces of the femur and tibia, the bones were cut into approximately 1 mm long fragments and digested with 1 mg/ml collagenase I/dispase II (Roche, 46793022). After incubating for 40 min at 37°C, the cell suspension was filtered through a 40-μm cell strainer (Corning, 431751) to obtain a single cell suspension and centrifuged at 1500 × g for 5 min. To remove the red blood cells, cells were resuspended in 1x RBC lysis buffer (BioLegend, 420301) on ice for 5 min. The solution was then centrifuged at 500 × g for 5 min and resuspended in PBS. The sample was stained with trypan blue (Sigma), and cell survival was evaluated microscopically. Single cell suspension was loaded onto a microfluidic chip and single cell RNA-seq libraries were constructed according to the manufacturer's instructions (Singleron GEXSCOPE Single Cell RNAseq Library Kit, Singleron Biotechnologies). scRNA_seq