Общество с ограниченной ответственностью "Экселлент Кволити Брендинг" (RU)
发明人:
Козлов Глеб Владимирович (RU)
申请号：
RU2013150642/10
公开号：
RU2013150642A
申请日:
2013.11.14
申请国别(地区):
RU
年份:
2015
代理人:
摘要：
1. A method for isolating cells of fibroblast, comprising the steps that produce fence skin tissue biopsy obtained was washed in physiological saline supplemented with human albumin, washed biopsy treated dispase solution in the culture medium Dulbeccos Modified Dulbecco from glucose 1 g / l (DMEM -LG) to separation film epidermis from the dermis, the dermis resulting washed with DMEM-LG and milled mechanically, dermis fragments distributed over the petri dish and poured DMEM-LG, supplemented with fetal bovine serum (EFV), a solution of essential amino acids (MEM NEAA) and antibiotics, cultured cells are periodically replacing the culture medium after the fibroblasts migrate out of fragments and fill the entire surface of the bottom of the cup, they are removed by the enzyme, and washed with culture medium, the enzyme is passed through the filter and pelleted by centrifugation, the cell pellet was resuspended in growth medium and plated onto culture flasks, placed in a CO-incubator and after reaching the confluent cell monolayer is removed by the enzyme, was precipitated and passaged until q.s. dimogo therapeutic kolichestva.2. A method according to claim 1, characterized in that the fence skin tissue is performed in a volume of not more than 2 mm / 1 mm / 1 mm.3. A method according to claim 1, characterized in that the resultant fence biopsy was washed in saline, human albumin at a concentration of 1% .4 latter. A method according to claim 1, characterized in that the processing used to dispase solution has a concentration of 0.1% .5. A method according to claim 1, characterized in that the treatment biopsy dispase solution in DMEM-LG was carried out for 3 hours on the orbital shaker at constant dormancy1. Способ выделения клеток фибробластов, заключающийся в том, что производят забор тканей кожи, полученный биоптат промывают в физиологическом растворе, дополненном человеческим альбумином, промытый биоптат подвергают обработке раствором диспазы в культуральной сред
