Method for constructing tissue-engineering bone by pouring stem cell of inducing bone marrow ground substance into vivo
A technology of tissue engineering bone and stem cells, applied in prosthetics, bone implants, medical science, etc., can solve the limitations of clinical application, the difficulty of clinical application in the effectiveness and safety of gene therapy, and the adverse effects of cells on the coordinated release mechanism of multiple factors, etc. problems, to achieve the effect of promoting rapid vascularization, promoting the speed and quality of osteogenesis
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[0021] Taking this method to repair the large segmental bone defect of goat tibial shaft as an example
[0022] 1. Cultivation and expansion of goat BMSCs: extract goat autologous bone marrow respectively, use 1% pentobarbital sodium 0.3ml / kg intravenous anesthesia, sumaamp, atropine intramuscular injection compound anesthesia. After skin preparation at the bilateral iliac crests, 10ml of red bone marrow was extracted by puncturing with a 16-gauge bone needle under strict aseptic conditions. 2:1 serum-free medium, centrifuge at 1000r / min for 10min, discard the supernatant, wash 2-3 times with PBS, remove the upper layer of fat and tissue fluid, replace the lower cell layer (mixture containing BMSCs) with complete medium Resuspend, inhale into a petri dish, add complete medium, and culture in a 37°C, 5% CO2, saturated humidity incubator. Cultivate for about 2-3w, confluent into a monolayer, digest with a mixture of 0.25% trypsin and 0.02% EDTA, terminate the digestion, then pa...
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