Tissue engineering bone tissue and preparation method thereof
A tissue engineering bone and tissue engineering technology, applied in the direction of bone implants, etc., can solve the problems of lack of basic mechanical stiffness, lack of bone mineral, lack of basic mechanical strength, etc., to achieve good biodegradability and physical and mechanical properties, The effect of good 3D connectivity and good biocompatibility
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[0031] 1) In vitro isolation and culture of bone marrow mesenchymal stem cells:
[0032] 10ml of the patient's autologous bone marrow was extracted, separated, and nucleated cells were extracted, washed twice with PBS+EDTA. Place in fibronectin-coated culture dishes, add DMEM culture medium, and incubator at 37°C containing 5% carbon dioxide. Two days later, unattached cells were removed and cultured in DMEM medium.
[0033] Subculture: When the cells are confluent to 60%-75% density, rinse with calcium and magnesium-free PBS, digest with trypsin, add DMEM medium containing fetal bovine serum to stop digestion, transfer to a centrifuge tube, and centrifuge. Discard the supernatant, add DMEM culture medium containing DMEM bovine serum, mix well, count, and subculture at a ratio of 1:2. Change the culture medium every 2-3 days, and subculture again when the cells reach a density of 60%-75%.
[0034] 2) Bone marrow mesenchymal stem cells are inoculated on cancellous bone scaff...
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