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СПОСОБ ВОССТАНОВЛЕНИЯ КОСТНЫХ ДЕФЕКТОВ ТРУБЧАТЫХ КОСТЕЙ КРИТИЧЕСКОЙ ВЕЛИЧИНЫ
专利权人:
Тепляшин Александр Сергеевич (RU)
发明人:
Тепляшин Александр Сергеевич (RU)
申请号:
RU2013140871/15
公开号:
RU2013140871A
申请日:
2013.09.05
申请国别(地区):
RU
年份:
2015
代理人:
摘要:
1. A process for preparing graft for bone regeneration tubular bones, comprising the steps that take aspirate bone marrow derived bone marrow samples were collected in tubes containing Li-heparin was then diluted with phosphate-buffered saline, filtered through a filter with a pore size of 70 microns, cells were centrifuged at 400 g for 10 minutes to obtain a precipitate, the resulting cell suspension was seeded into flasks of 150 smiz rate of 1 × 10kletok / cm, mesenchymal stem cells are cultured in DMEM with low glucose, complement ennoy 10% fetal calf serum, nonessential amino acid solution and antibiotics, the cultivation is carried out at a temperature of 37 ° C and 5% COv air, changing media every 3 days after reaching confluency the monolayer cells were removed from the substrate solution with 0.05% trypsin-EDTA and seeded culture flasks at the rate of 1 × 10kletok / CMB growth medium immediately before graft preparation procedure cells are removed from the substrate, washed free DMEM medium, resuspended in physiological saline, then the cell suspension was applied slowly on the matrix was incubated 3 hours at 37 ° C with shaking on an orbital shaker at a speed of 25 revolutions per minute, thereafter, the matrix is ​​poured DMEM medium supplemented SEC (fetal bovine serum), 10% dexamethasone 10M, β-glycerophosphate 10 mM, ascorbate - 2 phosphate 0.05 mM, 1% antibiotic - streptomycin at a concentration of 100 .mu.g / ml penicillin and 100 U / ml, every third day, the medium was changed to fresh, culturing was carried out for 28 dney.2. A process for preparing graft for bone regeneration tubular bones, comprising the1. Способ получения трансплантата для регенерации костной ткани трубчатых костей, заключающийся в том, что берут аспират костного мозга, полученные пробы костного мозга собирают в пробирки с литий-гепарином, затем разбавляют фосфатно-солевым буфером, фильтруют через фильтр с размером пор 70 мкм, центрифугируют клетки при 400 g в течение 10 мин для
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