bone marrow cells, control
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 37C, the cell suspension was filtered through a 40-m 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 6 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 manufacturers instructions (Singleron GEXSCOPE Single Cell RNAseq Library Kit, Singleron Biotechnologies). scRNA_seq