研究肝臟功巨噬功能能時,我們經常用荷蘭liposoma的巨噬細胞清除劑Clodronate Liposomes氯膦酸二鈉脂質體(貨號CP-005-005)清除肝臟巨噬細胞KF、然后回輸修飾或者感興趣的目的巨噬細胞。既要確保原位的巨噬細胞被很好的清除,又要讓回輸的巨噬細胞在早起階段及時發(fā)揮作用且不能被巨噬細胞清除劑清除了。這種既要,又要,就要求注射清除劑后,合適的時間點回輸目的細胞。
肝臟巨噬細胞清除,建議尾靜脈注射。注射后,一般24h后,肝臟定居巨噬幾乎都被清除。48h就可以回輸目的細胞。鑒于回輸實驗比較難做,回輸細胞的數量,活率,純度以及功能性活性都決定了Transfer實驗的成功與否。
如下這張圖,是機體巨噬細胞清除后,目的巨噬細胞制備回輸再構的流程示意圖。
以常見的IR模型為例。對于細胞回輸,一般選擇尾靜脈注射細胞,但是對于缺血再灌注模型(IR),可以考慮脾臟內注射。
Macrophage adoptive transfer缺血再灌注模型,回輸巨噬細胞
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.