文献类型：专利

英文题名：Dynamic suspension culture of cytokine-induced killer cell by isolating blood mononuclear cells, inoculating cells in medium, culturing with cytokine combination for stimulation and subjecting to dynamic suspension culture

作者：CAI H;ZHANG W;TAN W

机构：[1]UNIV EAST CHINA SCI & TECHNOLOGY

申请号：CN106754702-A

申请日：2016-12-26

公开日：2017-05-31

语种：英文

收录：DERWENT

摘要：NOVELTY - Dynamic suspension culture of cytokine-induced killer cell involves isolating blood mononuclear cells by density gradient centrifugation, inoculating freshly isolated blood mononuclear cells in complete Roswell Park Memorial Institute (RPMI) 1640 medium, culturing with cytokine combination for stimulation, subjecting to dynamic suspension culture, adjusting the addition of culture medium and cytokine combination in accordance to the cell density, readjusting the cell density and continuously culturing to obtain product. USE - Dynamic suspension culture of cytokine-induced killer cell (claimed). ADVANTAGE - The method improves cytokine-induced killer cell in vitro culture environment, adjusts the culture medium and the cytokine supplementation rate, reduces the influence of the culture environment on the cell growth, maintains the density of the cells during the culture and promotes the in vitro amplification effect and stability of the cytokine-induced killer cell. DETAILED DESCRIPTION - Dynamic suspension culture of cytokine-induced killer cell involves (a) isolating blood mononuclear cells by density gradient centrifugation, (b) inoculating 1-20x 105 cells/ml freshly isolated blood mononuclear cells in complete Roswell Park Memorial Institute (RPMI) 1640 medium and culturing the isolated blood mononuclear cells with cytokine combination for stimulation, (c) subjecting to dynamic suspension culture at 37 degrees C and 50-130 revolutions per minute (rpm) in a culture device in a shaker of 5% carbon dioxide incubator and (d) adjusting the addition of culture medium and cytokine combination in accordance to the cell density, gradually increasing the working volume of culture medium, maintaining the culture volume at 8-40% with respect to the total volume of the incubator, sampling and counting the cells everyday, calculating the cell density, adding the complete RPMI1640 medium containing 500 international units (IU)/ml interleukin-2 (IL-2) in accordance to the cell density, readjusting the cell density to 1-20x 105 cells/ml and continuously culturing for 14 days to obtain product.