Tissue-engineered alveolar bone tissue and preparation method thereof
A tissue engineering, alveolar bone technology, applied in biochemical equipment and methods, tissue regeneration, tissue culture and other directions, to achieve the effects of reducing medical costs, convenient material acquisition, and easy access
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Embodiment 1
[0036] Bone marrow stromal stem cells are used as seed cells for tissue engineered alveolar bone. Now, bone marrow stromal stem cells are isolated, cultured and expanded to prepare bone marrow stromal stem cell aggregates, and then bone marrow stromal stem cell aggregates are combined with scaffold materials in vitro to form tissue engineered alveolar bone tissue. , the specific preparation steps are as follows:
[0037] Step 1. Isolation, cultivation and screening of bone marrow mesenchymal stem cells: extract 3mL from the ilium, inject into a centrifuge tube containing 0.3mL heparin sodium saline (1000 U / mL), shake and then add 6mL lymphocyte separation solution (1.2 g / mL), remove the white membranous layer of mononuclear cells after centrifugation, wash twice with PBS, discard the supernatant after centrifugation, add stem cell medium to resuspend, and place in 5% CO 2 , Cultured at 37°C, changing the medium every other day, passaged when the cells grew to 80%, and labeled ...
Embodiment 2
[0045] The seed cells of tissue-engineered alveolar bone adopt periodontal ligament stem cells, which are now separated, cultured and expanded to prepare periodontal ligament stem cell aggregates, and then the periodontal ligament stem cell aggregates are combined with scaffold materials in vitro to form tissue-engineered teeth. Groove bone tissue, the specific preparation steps are as follows:
[0046] Step 1. Isolation, culture and screening of periodontal ligament stem cells: Collect 1 / 3 periodontal ligament tissue from the root of the third molar, cut it into pieces, and digest it with 0.2g / L type I collagenase and 0.4g / L Dispase enzyme for 1.5 hours Until the tissue is loose, sieve to obtain a single cell suspension, discard the supernatant after centrifugation, add stem cell medium to resuspend, and place in 5% CO 2 、Cultivate at 37°C, change the medium once every other day, pass passage when the cells grow to 75%, and label the first-generation periodontal ligament stem...
Embodiment 3
[0054] The seed cells of tissue-engineered alveolar bone adopt adipose stem cells, which are now separated, cultured and expanded to prepare adipose-derived stem cell aggregates, and then compound the adipose-derived stem cell aggregates with scaffold materials in vitro to form tissue-engineered alveolar bone tissue. Specific steps are as follows:
[0055] Step 1. Extract 3-10 mL of adipose tissue from the buccal fat pad of the oral cavity, put it in PBS containing penicillin and streptomycin 100 U / mL, soak it in a petri dish, wash it 3 times, each time for 4 minutes, and cut the adipose tissue into small pieces. 1mm 3 , add 0.25% type I collagenase 3 times the volume of adipose tissue, shake, digest at 37°C for 1 hour, add medium to stop digestion. Use 200μm and 100μm filters to filter in turn, centrifuge to collect the lower stromal vascular cell mixture (SVF), resuspend in PBS, filter with a 70μm filter, discard the supernatant after centrifugation, add medium to resuspend...
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