两种免疫磁珠分离法分选小鼠骨髓粒-单核祖细胞的效率比较

Isolation and Purification of Murine Granulocyte-monocyte Progenitor with Two Different Magnetic Activated Cell Sorting Methods

目的:提取小鼠骨髓细胞(bone marrow cell,BMC),用两种不同的免疫磁珠分离(magnetic activated cell sorting,MACS)试剂盒从小鼠BMC中分选提纯粒-单核祖细胞(granulocyte-monocyte progenitor,GMP),比较这两种免疫磁珠的分选效率。方法:从小鼠股骨和胫骨中提取BMC,通过两种不同的MACS试剂盒,即Lineage阳性细胞清除试剂盒和CD117阳性细胞分选试剂盒,分别得到Lineage-细胞群和CD117+细胞群,用代表GMP细胞表面标志物的荧光抗体标记,孵育后通过流式细胞荧光分选技术得到GMP细胞,并且对比得到GMP细胞的效率。结果:每2只野生型C57BL/6J小鼠可共收集骨髓细胞(7.02±1.24)×10^(7)个,细胞活力为(91.86±5.24)%。经过Lineage阳性细胞清除试剂盒得到的细胞数量为(5.71±2.86)×10^(6)个;经过CD117阳性细胞分选试剂盒得到的细胞数量为(2.70±0.56)×10^(6)个。Lineage磁珠分选纯化得到的GMP细胞数占总细胞数的比例为(10.90±1.37)%,CD117磁珠分选纯化得到的GMP细胞数占总细胞数的比例为(4.83±2.08)%。结论:Lineage阳性细胞清除试剂盒能更有效分选小鼠骨髓细胞中的粒-单核祖细胞。

Objective:Two different magnetic activated cell sorting(MACS)kits were used to sort and purify granulocyte-monocyte progenitor(GMP)cells from mouse bone marrow cells(BMC),comparing their sorting efficiency.Methods:BMC were extracted from mouse femur and tibia,and Lineage-cell population and CD117+cell population were obtained with two different MACS kits,Lineage cell depletion Kit and CD117 MicroBeads,respectively.Acquired cell population were stained and sorted by flow cytometry to obtain GMP cells,and the percentage of GMP cells was compared.Results:A total of(7.02±1.24)×10^(7)BMC cells were collected from each 2 wild-type C57BL/6J mice,and the cell viability was(91.86±5.24)%.The number of cells obtained by Lineage positive cell removal kit was(5.71±2.86)×10^(6),while the number of cells obtained by CD117 positive cell sorting kit was(2.70±0.56)×10^(6).The ratio of the number of GMP cells to total cells obtained by Lineage cell depletion Kit was(10.90±1.37)%,and the ratio of GMP cells obtained by CD117 MicroBeads was(4.83±2.08)%.Conclusions:The Lineage cell depletion Kit can more effectively sort granulocyte-mononuclear progenitor cells from mouse bone marrow cells.