肝臟巨噬細(xì)胞清除后回輸巨噬細(xì)胞方案和流程

研究肝臟功巨噬功能能時(shí)，我們經(jīng)常用荷蘭liposoma的巨噬細(xì)胞清除劑Clodronate Liposomes氯膦酸二鈉脂質(zhì)體（貨號(hào)CP-005-005）清除肝臟巨噬細(xì)胞KF、然后回輸修飾或者感興趣的目的巨噬細(xì)胞。既要確保原位的巨噬細(xì)胞被很好的清除，又要讓回輸?shù)木奘杉?xì)胞在早起階段及時(shí)發(fā)揮作用且不能被巨噬細(xì)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(shí)間點(diǎn)回輸目的細(xì)胞。

肝臟巨噬細(xì)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(xì)胞。鑒于回輸實(shí)驗(yàn)比較難做，回輸細(xì)胞的數(shù)量，活率，純度以及功能性活性都決定了Transfer實(shí)驗(yàn)的成功與否。

如下這張圖，是機(jī)體巨噬細(xì)胞清除后，目的巨噬細(xì)胞制備回輸再構(gòu)的流程示意圖。

以常見的IR模型為例。對(duì)于細(xì)胞回輸，一般選擇尾靜脈注射細(xì)胞，但是對(duì)于缺血再灌注模型（IR），可以考慮脾臟內(nèi)注射。

Macrophage adoptive transfer缺血再灌注模型，回輸巨噬細(xì)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.