巨噬細(xì)胞清除劑Clodronate Liposome是(Liposoma高端化,CP-005-005)是一種可以利用巨噬細(xì)胞的內(nèi)吞機(jī)制,將氯膦酸鹽帶入細(xì)胞內(nèi)充足,誘導(dǎo)巨噬細(xì)胞凋亡的試劑深入開展。它是由荷蘭阿姆斯特丹Vrije University 的Nico van Rooijen教授開發(fā)的更為一致,也叫做Clodronate Liposomes(氯膦酸二鈉脂質(zhì)體)。它是一種成熟技術的開發,方便研究與應用,經(jīng)濟(jì)的巨噬細(xì)胞清除方法,可以有效地清除動物體內(nèi)不同組織部位和血液中的巨噬細(xì)胞更高效。它是研究巨噬細(xì)胞功能的重要手段全面協議。荷蘭Liposoma巨噬細(xì)胞清除劑頻繁見刊于Cell,Nature和Science具體而言。
我們在清除巨噬細(xì)胞后工具,經(jīng)常想回輸我們修飾或者刺激或者目的巨噬細(xì)胞。如何做好回輸實(shí)驗(yàn)喜愛?如何確保清除體內(nèi)巨噬細(xì)胞而又不會清除回輸巨噬細(xì)胞重要的角色?把握好時間點(diǎn)和參考類似的文獻(xiàn)至關(guān)重要。這篇JEM高質量,模型是OVA誘導(dǎo)的哮喘模型也逐步提升,關(guān)注的巨噬細(xì)胞是肺泡巨噬細(xì)胞。這篇JEM就是用的荷蘭Liposoma的巨噬細(xì)胞清除劑智能設備,然后回輸目的巨噬細(xì)胞。
荷蘭Liposoma的巨噬細(xì)胞清除劑清除方案和回輸方案:
給藥方式:intratracheally (i.t.) 氣管給藥
劑量:40ul
次數(shù):2次
時間點(diǎn):造模后第18天和第19積極回應,連續(xù)兩天
回輸細(xì)胞數(shù)量:5*10^5(活率擴大公共數據,活性和活化這些都需要評估)
回輸時間點(diǎn):最后一次給巨噬細(xì)胞清除劑后間隔24h
品牌:Liposoma
貨號:CP-005-005
規(guī)格:5ml+5ml
品名:Clodronate Liposomes and PBS Liposomes
Adoptive transfer of AMs was performed as previously reported (Miki et al., 2021; Qian et al., 2015). For in vivo deletion of macrophages in lung tissues, mice were sensitized with OVA as described above and treated with 40 μl of clodronate liposome i.t. for two successive days (days 18 and 19). For the AM adoptive transfer study, AMs derived from WT or AMFR knockout mice were then transferred by i.t. injection into the lungs of clodronate liposome-treated and OVA-sensitized WT mice at a density of 5 × 105 cells/mouse (40 μl) on day 20. 24 h after AM delivery, the mice were i.t. challenged with OVA for three days (days 21, 22, and 23). On day 25, the mice were sacrificed to analyze allergic asthmatic inflammation (如上圖).
Similarly, for in vivo knockdown of CIS in mice lung macrophages, OVA-sensitized recipient LysMcreAmfrfl/fl mice with clodronate liposome i.t. treatment received i.t. Amfr?/? BMDMs that were infected with either Scr shRNA or Cish shRNA at a density of 5 × 105 cells/mouse (40 μl) on day 20. 24 h after macrophage delivery, the mice were i.t. challenged with OVA for 3 d (days 21, 22, and 23). On day 25, the mice were sacrificed to analyze allergic asthmatic inflammation (見下圖).
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