Preparation of cell transplant
A technology of grafts and cells, applied in the field of preparation of "allogeneic cells-preparation-tissue engineering" grafts, which can solve the problems of rejection that have not been mentioned
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[0046] Materials and methods
[0047] MSC Isolation, Characterization and Culture
[0048] MSCs were obtained from two inbred strains of mice: C57BL / 6J(B6) and Balb / c. Male mice aged 4-6 weeks were sacrificed by cervical dislocation. The femur and tibia were removed, cleaned of all connective tissue, and placed in 5 mL of α-MEM supplemented with 1× penicillin / streptomycin on ice. All tibia and femur ends were cut off to expose the bone marrow, inserted into the prepared centrifuge tube, and centrifuged at 400g for 1 minute to collect the bone marrow. Resuspend the precipitate in 3 mL of complete medium (CM: α-MEM supplemented with 16.6% fetal bovine serum (FBS), 100 U / mL penicillin, 100 g / mL streptomycin), and use a 21-gauge injection needle to pass through 70-μm nylon mesh for filtration. Cells from each long bone were placed in 10 cm dishes containing 40 mL of CM. After 24 hours, remove unattached cells with phosphate-buffered saline (PBS), and then add 10 mL of fresh C...
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