研究肝脏功巨噬功能能时,我们经常用荷兰liposoma的巨噬细胞清除剂Clodronate Liposomes氯膦酸二钠脂质体(货号CP-005-005)清除肝脏巨噬细胞KF、然后回输修饰或者感兴趣的目的巨噬细胞。既要确保原位的巨噬细胞被很好的清除,又要让回输的巨噬细胞在早起阶段及时发挥作用且不能被巨噬细胞清除剂清除了。这种既要,又要,就要求注射清除剂后,合适的时间点回输目的细胞。
肝脏巨噬细胞清除,建议尾静脉注射。注射后,一般24h后,肝脏定居巨噬几乎都被清除。48h就可以回输目的细胞。鉴于回输实验比较难做,回输细胞的数量,活率,纯度以及功能性活性都决定了Transfer实验的成功与否。
如下这张图,是机体巨噬细胞清除后,目的巨噬细胞制备回输再构的流程示意图。
以常见的IR模型为例。对于细胞回输,一般选择尾静脉注射细胞,但是对于缺血再灌注模型(IR),可以考虑脾脏内注射。
Macrophage adoptive transfer缺血再灌注模型,回输巨噬细胞
Mice were injected i.v. with 200 μl clodronate liposomes (Liposoma, Netherlands) 48 h before transfer for macrophage depletion. Then, the mouse partial liver IRI model was established in recipient mice. Freshly isolated intrahepatic macrophages from young or aged C57BL/6 mice were transplanted into aged or young recipient mice by intrasplenic injection (using a 30-G insulin syringe) at the time of reperfusion.