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METHOD FOR REPAIRING LONG BONE DEFECTS OF CRITICAL SIZE
METHOD FOR REPAIRING LONG BONE DEFECTS OF CRITICAL SIZE
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机译:修复严重骨长骨缺损的方法
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FIELD: medicine.;SUBSTANCE: group of inventions refers to medicine, namely to orthopaedics, and can be used for producing a transplant for long bone tissue repair. That is ensured by collecting bone marrow aspirate into lithium heparin test tubes, diluting with phosphate salt buffer, filtered through a filter with pore size 70 mcm and centrifuging for 10 min at 400 g. That is followed by inoculating flasks 150 cm2 with the produced mesenchymal stem cells (MSCs) at 1×106 cells/cm2, culturing at temperature 37°C and 5% CO2 in air, in the Dulbecco's Modified Eagle's medium containing glucose 1 g/l and less, with 10% embryo calve serum. The medium is changed every 3 days. Before preparing the transplant, the cells are removed from the substrate with 0.05% trypsin EDTA, washed and re-suspended. The cell suspension is applied on the matrix, incubated for 3 hours at temperature 37°C with rocking in a rotation shaker at 25 rpm. The matrix is filled with the DMEM medium with 10% bovine foetal serum, dexamethasone 10-7 M, β-glycerophosphate 10 mM, ascorbate-2-phosphate 0.05 mM, 1% streptomycin 100 mcg/ml or penicillin 100 units/ml. The MSCs are cultured for 28 days. What is also presented is a method for long bone tissue repair.;EFFECT: group of inventions provides the uniform bone tissue repair in replacing the bone tissue defect of critical size with the transplant.;3 cl, 2 tbl, 6 ex
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机译:领域:发明领域:一组发明涉及医学,即骨科,并且可以用于生产用于长骨组织修复的移植物。通过将骨髓抽吸液收集到肝素锂试管中,用磷酸盐缓冲液稀释,通过孔径为70 mcm的过滤器过滤,并在400 g下离心10分钟来确保这一点。随后,以产生的间充质干细胞(MSCs)以1×10 6 Sup> cells / cm 2 Sup>接种150 cm 2 Sup>的烧瓶,进行培养在Dulbecco改良的Eagle's培养基(温度为37°C,空气中含5%CO 2 Sub>)中,葡萄糖含量为1 g / l和更少,并有10%的犊牛血清。介质每3天更换一次。在准备移植之前,先用0.05%胰蛋白酶EDTA从底物中除去细胞,然后洗涤并重悬。将细胞悬浮液施加到基质上,在旋转摇床上以25 rpm摇动,在温度37°C下孵育3小时。基质填充有DMEM培养基,其中含有10%的牛血清,地塞米松10 -7 Sup> M,β-甘油磷酸10 mM,抗坏血酸-2-磷酸酯0.05 mM,1%链霉素100 mcg / ml或青霉素100单位/毫升。将MSC培养28天。还提出了一种用于长骨组织修复的方法。效果:一组发明提供了均匀的骨组织修复,以用移植物替代临界大小的骨组织缺损。3cl,2 tbl,6 ex
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