流式细胞术分选小鼠骨髓造血干细胞的技术方案  

Technical scheme of sorting mouse bone marrow hematopoietic stem cells by flow cytometry

作　　者：户乃丽 Hu Nai-li(Central Laboratory,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学中心实验室,北京100069

出　　处：《分析仪器》2022年第5期61-65,共5页Analytical Instrumentation

摘　　要：流式细胞术是检测和分选造血干细胞的重要手段,根据其表型和功能,目前有多种不同组合的表面标记物用于识别小鼠的骨髓造血干细胞。但是由于其含量极低,因此在实验操作中存在着一定难度。本实验以目前公认度较高的LSK方案结合SLAM家族分离策略为依据,首先对小鼠骨髓细胞分群及亚型检测,而后结合生物素标记的免疫磁珠对样品进行预纯化,再经流式分选,得到了纯度和活性均高于90%的LSK阳性造血干细胞。本文详细介绍了染色的方案及具体操作,以及流式分选的技术细节。Flow cytometry is an important method for detecting and sorting hematopoietic stem cells.There are many different combinations of surface markers used to identify mouse bone marrow hematopoietic stem cells.However,due to the very low content of bone marrow hematopoietic stem cells,it is difficult to operate in practice.The dyeing scheme and specific operation,as well as the technical details of flow sorting are introduced in this paper.Based on the recognized LSK scheme combined with the SLAM family separation strategy,the subtypes and population of mouse bone marrow cells are detected firstly,then the samples are pre purified by biotin-labeled immunomagnetic beads.Finally,LSK positive hematopoietic stem cells with purity and activity higher than 90%are obtained by flow cytometric analysis.

关 键 词：造血干细胞 免疫磁珠分选 流式分选 

分 类 号：R-33[医药卫生]