Bone tissue constructed by tissue engineering
A tissue engineering and bone tissue technology, applied in the field of biomedical engineering, can solve the problems of lack of bone mineral, lack of basic mechanical stiffness, lack of basic mechanical strength, etc., and achieve the effect of good biocompatibility
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[0027] l) In vitro isolation and culture of bone marrow mesenchymal stem cells: extract 5-10 ml of the patient's own bone marrow, separate with Ficoll, extract nucleated cells, and wash twice with PBS + EDTA. Add DMEM (purchased from GIBCOBRL company. Contains 20% fetal bovine serum) culture medium, place in fibronectin-coated culture dish, 37 ℃ containing 5% carbon dioxide incubator. After two days, unattached cells were removed and cultured in conditioned medium.
[0028] Subculture
[0029] When the cells are confluent to 60%-75% density, rinse with 10ml of calcium and magnesium-free PBS, digest with 0.25% trypsin at 37°C for 5 minutes, add DMEM culture solution containing 10% fetal bovine serum to stop the digestion, transfer to a centrifuge tube, Centrifuge at 1200r / min for 5 minutes. Discard the supernatant, add 10ml of DMEM culture solution containing 10% fetal bovine serum, mix well, count, and subculture at a ratio of 1:3. Change the medium every 2-3 days until the...
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