Construction and application of a coaxial vascularized tissue engineered bone
A tissue-engineered bone and vascularization technology, used in tissue regeneration, femur, bone implants, etc., can solve problems such as failure of large-volume tissue-engineered bone construction, failure to achieve early vascularization in the axial area, etc., to promote osteogenesis. effect of effect
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Embodiment 1
[0031] Example 1: Experimental study on in vivo rapid formation of vascular network in hydrogel loaded with vascularized seed cells
[0032] (1) Acquisition of seed cells:
[0033]1) Acquisition of bone marrow endothelial progenitor cells (EPCs): extract 5ml of bone marrow, obtain mononuclear cells in the bone marrow by gradient centrifugation, culture in endothelial cell culture medium, change the medium every 2-3 days, and cultivate for 2-4 weeks Finally, through morphological observation and flow cytometry detection to identify the cell properties, after counting, they were inoculated into culture flasks for subculture, and the 3rd generation cells were taken for future use (see figure 2 );
[0034] 2) Acquisition of bone marrow mesenchymal stem cells (BMSCs): According to the bone marrow culture method, extract red bone marrow, anticoagulate with 500u / mL heparin solution, mix well, centrifuge at 1000r / min for 10min, discard the supernatant, add DMEM complete medium, reco...
Embodiment 2
[0041] Example 2: Experimental study of coaxial tissue engineered bone repairing load-bearing bone defects
[0042] (1) Acquisition of seed cells:
[0043] 1) Acquisition of bone marrow endothelial progenitor cells (EPCs): extract 5ml of bone marrow, obtain mononuclear cells in the bone marrow by gradient centrifugation, culture in endothelial cell culture medium, change the medium every 2-3 days, and cultivate for 2-4 weeks Afterwards, through morphological observation and flow cytometry detection to identify the nature of the cells, after counting, they were inoculated into culture flasks for subculture, and the 3rd generation cells were taken for later use;
[0044] 2) Acquisition of bone marrow mesenchymal stem cells (BMSCs): According to the bone marrow culture method, extract red bone marrow, anticoagulate with 500u / mL heparin solution, mix well, centrifuge at 1000r / min for 10min, discard the supernatant, add DMEM complete medium, record After a few days, they were inoc...
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