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巨噬細(xì)胞清除劑清除肺泡巨噬細(xì)胞后Transfer回輸實(shí)驗(yàn)解決方案
巨噬細(xì)胞清除劑Clodronate Liposome是（Liposoma，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)，誘導(dǎo)巨噬細(xì)胞凋亡的試劑。它是由荷蘭阿姆斯特丹Vrije University 的Nico van Rooijen教授開發(fā)的，也叫做Clodronate Liposomes（氯膦酸二鈉脂質(zhì)體）。它是一種成熟，方便，經(jīng)濟(jì)的巨噬細(xì)胞清除方法，可以有效地清除動(dòng)物體內(nèi)不同組織部位和血液中的巨噬細(xì)胞。它是研究巨噬細(xì)胞功能的重要手段。荷蘭Liposoma巨噬細(xì)胞清除劑頻繁見刊于Cell，Nature和Science。
我們?cè)谇宄奘杉?xì)胞后，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)？如何確保清除體內(nèi)巨噬細(xì)胞而又不會(huì)清除回輸巨噬細(xì)胞？把握好時(shí)間點(diǎn)和參考類似的文獻(xiàn)至關(guān)重要。這篇JEM，模型是OVA誘導(dǎo)的哮喘模型，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，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(shí)間點(diǎn)：造模后第18天和第19，連續(xù)兩天
回輸細(xì)胞數(shù)量：5*10^5(活率，活性和活化這些都需要評(píng)估)
回輸時(shí)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(hào)：CP-005-005
規(guī)格：5ml+5ml
品名：Clodronate Liposomes and PBS Liposomes
AM adoptive transfer
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).
CIS knockdown in vivo
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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