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.