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巨噬细胞清除剂清除肺泡巨噬细胞后Transfer回输实验解决方案

更新时间:2024-11-20   点击次数:159次

巨噬细胞清除剂Clodronate Liposome是(Liposoma,CP-005-005)是一种可以利用巨噬细胞的内吞机制,将氯膦酸盐带入细胞内,诱导巨噬细胞凋亡的试剂它是由荷兰阿姆斯特丹Vrije University 的Nico van Rooijen教授开发的,也叫做Clodronate Liposomes(氯膦酸二钠脂质体)它是一种成熟,方便,经济的巨噬细胞清除方法,可以有效地清除动物体内不同组织部位和血液中的巨噬细胞它是研究巨噬细胞功能的重要手段。荷兰Liposoma巨噬细胞清除剂频繁见刊于Cell,Nature和Science。

我们在清除巨噬细胞后,经常想回输我们修饰或者刺激或者目的巨噬细胞。如何做好回输实验?如何确保清除体内巨噬细胞而又不会清除回输巨噬细胞?把握好时间点和参考类似的文献至关重要。这篇JEM,模型是OVA诱导的哮喘模型,关注的巨噬细胞是肺泡巨噬细胞。这篇JEM就是用的荷兰Liposoma的巨噬细胞清除剂,然后回输目的巨噬细胞。



荷兰Liposoma的巨噬细胞清除剂清除方案和回输方案:

给药方式:intratracheally (i.t.) 气管给药

剂量:40ul

次数:2次

时间点:造模后第18天和第19,连续两天

回输细胞数量:5*10^5(活率,活性和活化这些都需要评估)

回输时间点:最后一次给巨噬细胞清除剂后间隔24h

品牌:Liposoma

货号:CP-005-005

规格:5ml+5ml

品名:Clodronate Liposomes and PBS Liposomes


巨噬细胞清除剂清除肺泡巨噬细胞后Transfer回输实验解决方案


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 (如上图).

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 (见下图).

巨噬细胞清除剂清除肺泡巨噬细胞后Transfer回输实验解决方案




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