免疫磁珠法分选小鼠肝脏Kupffer细胞及其特性观察

目的:建立免疫磁珠法,分离纯化小鼠肝脏Kupffer细胞并进行功能鉴定。方法:采用原位肝脏酶灌注、制备肝非实质细胞悬液、加入PE标记的大鼠抗小鼠F4/80一抗,然后结合山羊抗大鼠IgG Dynabeads免疫磁珠分选Kupffer细胞,流式细胞仪检测其纯度并检测其吞噬功能。结果:在经过胶原酶Ⅳ型消化、免疫磁分选,获得小鼠肝Kupffer细胞产量约为(8.05±0.34)×106/鼠肝,细胞纯度为(96.6±0.7)%,细胞活力大于90%。结论:应用免疫磁珠法能有效地分离纯化小鼠肝脏Kupffer细胞,为体外进一步研究提供了高纯度和活力的Kupffer细胞群。

Isolation and purification of mouse liver Kupffer cells by immunomagnetic bead separation

Objective:To establish the way of immunomagnetic bead separation for isolating and purifying mouse liver Kupffer cells and exploring their phagocytic function.Methods:The techniques included the processes of liver perfusion in situ,preparation of non-parenchymal cells suspension,isolation using anti-mouse F4/80 antibody and sheep anti-rat IgG Dynabeads.Finally,flow cytometry was used to detect the purity and phagocytic function.Results:By collagenase Ⅳ digestion and immunomagnetic bead separation,Kupffer cells were obtained in the amount of(8.05±0.34)×106/mouse liver.The purity was(96.6±0.7)% and more than 90% of cells were viable.Conclusion:Mouse Kupffer cells can be isolated and purified by immunomagnetic bead separation,which makes it possible to further the study of this cell population in vitro.