Gosudarstvennoe byudzhetnoe uchrezhdenie zdravookhraneniya "Nauchno-issledovatelskij institut - Kraevaya klinicheskaya bolnitsa N 1 imeni professora S.V. Ochapovskogo" Ministerstva zdravookhraneniya K
发明人:
Gilevich Irina Valerievna (RU),Гилевич Ирина Валериевна (RU),Sotnichenko Aleksandr Sergeevich (RU),Сотниченко Александр Сергеевич (RU),Melkonyan Karina Igorevna (RU),Мелконян Карина Игоревна (RU),Yuts
申请号：
RU2019133278
公开号：
RU0002717088C1
申请日:
2019.10.18
申请国别(地区):
RU
年份:
2020
代理人:
摘要：
FIELD: medicine.SUBSTANCE: invention refers to medicine, namely to regenerative medicine, and can be used in reconstructive surgery to create a technology for obtaining and using bioengineering matrix as a graft for practical purposes. That is ensured by sampling pig's dermis 3 mm thick after preliminary removal of the epidermis by dermatome, then samples are frozen to temperature of -80 °C, after defrosting samples are poured with Trypsin-Versene solution and placed in shaker-incubator in mode of 100 rpm at 37 °C for 18 hours； at the second stage, the samples are placed on a rotating platform in mode of 170 rpm and subjected to sequential cyclic action of detergent solutions: 1 % Triton X100 solution for 2 hours and 4 % sodium deoxycholate in combination with 0.002 M Na2-EDTA for 2 hours, total number of treatment cycles is equal to 5. After each detergent samples are washed in deionised water for 10 minutes. At the third stage, the samples are treated with a solution of porcine pancreatic DNase I (2,000 units are dissolved in 200 ml of a phosphate buffer with calcium and magnesium) in a shaker incubator in mode of 100 rpm at 37 °C for 4 hours. Decellularization is terminated by exposure to 10 % chlorhexidine bigluconate in phosphate buffer for 24 hours with solution change every 6 hours. That is followed by confirming the quality of decellulyarisation by histological examination, molecular-biological analysis and culture methods.EFFECT: method enables reducing exposure time of decellularising solutions, reducing the level of residual DNA in tissue up to 60 ng/mg of tissue in a wet sample.1 clИзобретение относится к медицине, а именно к регенеративной медицине, и может быть использовано в реконструктивной хирургии для создания технологии получения и использования в практических целях биоинженерного матрикса в качестве трансплантата. Для этого осуществляют забор свиной дермы толщиной 3 мм после предварительного снятия эпидермиса дерматомом, далее образцы замораживаю
