摘要
为研究人外周血单个核细胞(PBMC)分离优化的条件,提高细胞的分离率与收集率,在一定程度上减少分离时间和原料,提高细胞获得率与纯度,从而满足工艺化、市场化研发免疫产品的要求,本实验从分离离心方面和洗涤离心方面列出正交表探索实验找出最佳条件。分离离心时,转速为主要影响因素,600g为最佳离心转速;洗涤离心时,分离液体积为主要影响因素,2倍分离液体积为最佳分离液体积。洗涤缓冲液的选材中,0.9%的生理盐水获得活细胞的浓度最高。基于实验结论,可通过调整优化来满足实验需求来适应实验室各条件,分离出更有活力、更多的PBMC。
In order to study the optimized conditions of PBMC separation and improve the separation rate and collection rate of cells, the separation time and raw materials can be reduced to some extent, and the cell acquisition rate and purity can be improved, so as to meet the requirements of technological and market research and development of immune products. In this experiment, orthogonal tables were listed from the aspects of separation centrifuge and washing centrifuge to explore the best conditions. The speed of centrifugation is the main influencing factor, and 600g is the best centrifugal speed. When washing centrifuge, the volume of separated liquid is the main influencing factor, and 2 times the volume of separated liquid are the optimal conditions for washing collection. Among the selected materials of washing buffer, 0.9% normal saline obtained the highest concentration of living cells. Based on the experimental conclusions, the experimental requirements can be adjusted and optimized to adapt to the laboratory conditions, and more dynamic and more PBMC can be isolated.
作者
李飞
潘妍
张建平
Li Fei;Pan Yan;Zhang Jian-ping(Taizhou university, Zhejiang Taizhou 318000;Yeda Research Institute of Gene and Cell Therapy, Zhejiang Taizhou 318000)
出处
《生物化工》
2019年第5期68-70,共3页
Biological Chemical Engineering
关键词
外周血单个核细胞
离心力
密度梯度离心法
Peripheral blood mononuclear cell
Centrifugalforce
Density gradient centrifugation
