Fig. 1

Step-by-step process of isolating bone marrow endothelial cells: Day 1: a the mice were killed by cervical dislocations soak in 70% ethanol for 2 min. b The long bone of femora and tibias harvested was placed in a sterile 10-mm dish after dissecting the muscle, tendon, and ligaments. c The long bones were washed with DPBS/EDTA solution three times. d One end of the long bone cut-off with dissecting forceps inserted into the 0.5-ml Nest microtubes was centrifuge at 15,000 for 30 s. e The pellet of the bone marrow after centrifugation (f) The bone marrow pellet was suspended with DPBS/EDTA solution, filtered with a cell strainer, and centrifuged at 300g for 10 min at room temperature. Day 2: a the bone marrow cells resuspended with DMEM supplemented with 20% FBS plus antibiotics and incubated in the incubator overnight. Non-adherent cells were discarded, and marrow-adherent cells were washed twice with DPBS/EDTA solution. b Accutase solution poured into the dish to detach the attached cells for 15 min at room temperature. c The cells were resuspended with DPBS/EDTA solution and centrifuged at 300g for 10 min. d The resuspended bone marrow cells were incubated at 4°C with the CD31 microbeads. e The beads pass via the column and wash three times. f The magnetic cells resuspended with endothelial cell medium supplement with growth factors