seo 1.4
corning® nk 活化扩增培养kit， for activation and expansion of human natural killer cell
nk细胞活化扩增培养
自然杀伤细胞（nk）是重要的免疫细胞，能识别并清除肿瘤和病毒感染。体外大量培养nk细胞，满足免疫治疗所需一向比较困难。据资料，现有的培养方法都是采用扩增培养基搭配细胞因子（il-2，il-7，il-15）的方案，需要研究者自行优化后使用。而康宁的培养套装具备nk细胞活化扩增的全部材料及相应的标准操作程序，无需进一步的优化。
corning的nk培养套装包含抗体预包被t75细胞培养瓶，50ml nk细胞活化培养基，1.8ml nk细胞活化添加剂和1 l nk细胞扩增培养基。前三个组份特别设计用于nk细胞活化，而nk细胞扩增培养基用于nk细胞扩增。nk活化培养基和扩增培养基生产都是使用高质量试剂和cgmp级的原料。培养基唯二所含蛋白是注射级的血清白蛋白和重组人胰岛素。
corning® nk 活化扩增培养kit
英文名：corning® nk expansion kit
中文名：corning® nk细胞活化扩增培养套装
corning® nk 活化扩增培养kit组成
nk细胞活化培养基
nk细胞活化添加剂
nk细胞扩增培养基
抗体预包被t75细胞培养瓶
细胞培养袋
corning® nk 活化扩增培养kit特色
扩增能力强，在14天内可达到100倍扩增
效应细胞比例高，可达80％
操作方便
corning® nk 活化扩增培养kit订购信息
价格：12000元/套
品牌
货号
产品描述
包装
corning
88-570-kit
nk kit for activation and expansion of human natural killer cells
nk细胞活化/扩增培养基套装
1kit
套装包含
活化用
pre-coated t-75 flask 抗体预包被t75细胞培养瓶
1瓶
nk primary medium nk细胞活化培养基
1×50 ml
nk primary supplement nk细胞活化添加剂
1×1.8 ml
扩增用
expansion medium nk细胞扩增培养基
1×1000 ml
配件
细胞培养袋
1 x 640 cm2
上海华雅思创生物科技有限公司授权代理corning培养基，专业为生命科学领域内的科学研究、细胞治疗和生物制药相关人员提供优质产品和服务。
联系方式： 电话：021-64933729  手机：187 2127 8985  qq：1922853279
corning® nk kit活化扩增方法
1.  热灭活自体血浆，56°c，30min。离心， 800 xg ， 20 min，去除沉淀。4°c 保存上清血浆，培养备用。
2.  至少5倍体积的pbs（不含钙镁）清洗pbmcs。室温离心500xg，10min，收集pbmcs。 pbmcs稀释到 1 x 106 cells/ml，使用nk活化培养基配比1.8ml nk活化添加剂和10%自体血浆。
3. 预包被t-75培养瓶使用前，用pbs（不含钙镁）清洗两次。添加30 ml pbmc悬液到预包被t-75培养瓶，37°c，5% co2培养。
注意：第六天之前，如果培养基变黄，细胞密度超过2.0 x 106 cells/ml，需要添加新鲜的nk活化培养基配比10%自体血浆，保持细胞密度在在5 x 105到 2.0 x 106 cells/ml之间。
4.  培养第六天，培养液离心，适量nk扩增培养基（1000 iu/ml il-2 和10%自体血浆）重悬细胞，再转移到t-225培养瓶或透气细胞培养袋。
5. 每隔两三天，根据细胞扩增状态，培养体系中添加新鲜的nk扩增培养基（1000 iu/ml il-2 和10%自体血浆）保持细胞密度在5 x 105到 2.0 x 106 cells/ml之间。
6.  细胞总量到达2 x 109 后收获细胞用于cck8 杀伤力和免疫表型检测。
注意：整个实验过程中，细胞悬液吹打和培养容器拍打都要轻柔，以避免细胞损伤，保持细胞活性。
corning® nk expansion kit
natural killer (nk) cells are crucial immune cells that can recognize and kill tumors and virus infection. it is difficult to obtain the large numbers of nk cells ex vivo that are necessary for adoptive immunotherapy. several methods of nk activation/expansion have been reported that use expansion medium plus various cytokines (e.g., il-2, il-7, and il-15). however, these require further optimization by researchers.
the corning nk kit contains a pre-coated t-75 flask, 50 ml nk primary medium, 1.8 ml nk primary supplement, and 1l nk expansion medium. the first three components are specially designed for nk cell activation, whereas the nk expansion medium is utilized for nk cell expansion. both nk primary medium and nk expansion medium are manufactured using high quality reagents and cgmp-grade raw materials. the only proteins present in the media are injectable levels of serum albumin and recombinant human insulin.
nk cell activation and expansion
on day 0, the auto-plasma was inactivated by heating at 56°c for 30 min. and centrifuged at 800 xg for 20 min. to remove the precipitates. the supernatant was collected and stored at 4°c for further use.
the pbmcs were washed once with at least 5-fold pbs buffer, and the sample was centrifuged at 500 xg for 10 min. at room temperature. the density of pbmcs was adjusted to 1 x 106 cells/ ml using kbm nk primary medium containing 1.8 ml kbm nk primary supplement and 10% auto-plasma.
the pre-coated t-75 flask was washed carefully twice with pbs just before use. thirty milliliters of the suspension of pbmcs  were seeded into the pre-coated t-75 flask and incubated at 37°c in a humidified atmosphere of 5% co2 in air.
note: before day 6 when the medium turned yellow and the cell density was above 2 x 106 cells/ml, fresh kbm nk
primary medium plus 10% auto-plasma were added to the cell suspension to ensure that cell density stayed within the range of
2.5 x 105 to 2.0 x 106 cells/ml.
on day 6, all cells were centrifuged and re-suspended with an appropriate volume of kbm nk expansion medium  containing 1,000 iu/ml il-2 and 10% auto-plasma and then transferred to a new t-225 flask or gas-permeable culture bag.
in the following days, kbm nk expansion medium containing 1,000 iu/ml il-2 was added every 2 to 3 days based on the cell proliferation status to ensure that cell density stayed within
2.5 x 105 to 2.0 x 106 cells/ml.
the cells were harvested for cck8 cytotoxic and immuno- phenotyping testing when the total cell number exceeded 2 x 109.
note: the cells were gently dissociated, and the culture vessels were tapped softly to avoid cell damage and maintain cell viability throughout the entire experimental process.
上海华雅思创生物科技有限公司授权代理corning培养基，专业为生命科学领域内的科学研究、细胞治疗和生物制药相关人员提供优质产品和服务。
联系方式： 电话：021-64933729  手机：187 2127 8985  qq：1922853279

---

上海华雅思创生物科技有限公司

18721278985
中国 上海