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翌圣生物科技（上海）股份有限公司
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掃一掃訪問手機(jī)商鋪
巨噬細(xì)胞是體內(nèi)每個器官中都存在的細(xì)胞，存在于表皮、角膜和沒有血管的關(guān)節(jié)內(nèi)部，其體內(nèi)生物學(xué)研究的重要方法之一是巨噬細(xì)胞耗竭。清除巨噬細(xì)胞，可以全面了解巨噬細(xì)胞在病理?xiàng)l件下的功能。目前，巨噬細(xì)胞清除的主要方法包括構(gòu)建巨噬細(xì)胞缺失動物模型和使用藥物（氯膦酸鹽脂質(zhì)體）去除法，但是，構(gòu)建巨噬細(xì)胞缺失動物模型價(jià)格昂貴，周期長，因此，氯膦酸鹽脂質(zhì)體（Clodronate Liposomes）是目前最為成熟、方便、經(jīng)濟(jì)的一種巨噬細(xì)胞清除工具，可以有效的清除動物體內(nèi)包括肝臟，脾臟，肺部，血液等多個不同組織和部位中的巨噬細(xì)胞，也是目前使用泛的一種巨噬細(xì)胞清除的方法。
將氯膦酸鹽包裹進(jìn)磷脂體的水相中形成氯膦酸鹽脂質(zhì)體，氯膦酸鹽不能自由的通過脂質(zhì)體磷脂體雙分子層。注射到生物體內(nèi)后，氯膦酸鹽脂質(zhì)體會被巨噬細(xì)胞吞噬，在巨噬細(xì)胞內(nèi)的溶酶體磷酸酶的作用下，溶解在脂質(zhì)體內(nèi)的氯膦酸鹽會被逐步釋放出來并在細(xì)胞內(nèi)積累，當(dāng)達(dá)到一定濃度時(shí)，巨噬細(xì)胞將會受到不可逆損傷，誘發(fā)細(xì)胞凋亡。死亡細(xì)胞內(nèi)的氯膦酸鹽會被釋放到細(xì)胞外，隨尿液排出體外。
不同種類的小鼠給藥劑量略有差異，可根據(jù)以下表格中的給藥劑量摸索。
表1.C57小鼠巨噬細(xì)胞清除給藥方法
巨噬細(xì)胞有很多種亞型，不同的亞型部分標(biāo)志物不同（見表2），另外，不同組織和器官的巨噬細(xì)胞的標(biāo)志物也會有差異（見表3）。一般在做巨噬細(xì)胞清除實(shí)驗(yàn)前，應(yīng)先查找相關(guān)文獻(xiàn)，找出具體清除部位的巨噬細(xì)胞的表面標(biāo)志物，選擇最佳標(biāo)志物進(jìn)行流式實(shí)驗(yàn)。
表2.不同亞型的巨噬細(xì)胞標(biāo)志物
參考文獻(xiàn)：Klopfleisch R. Macrophage reaction against biomaterials in the mouse model - Phenotypes, functions and markers. Acta Biomater. 2016 Oct 1;43:3-13. doi: 10.1016/j.actbio.2016.07.003. Epub 2016 Jul 6. PMID: 27395828.
表3.不同組織中巨噬細(xì)胞的標(biāo)志物
小鼠品系：C57BL/6J小鼠
注射方式：腹腔注射
取樣部位：外周血
注射劑量：200 μl /只
抗體選擇：F4/80
檢測方法：巨噬細(xì)胞耗竭模型采用氯膦酸脂質(zhì)體，腹腔注射200μL每只小鼠，在注射氯鈉脂質(zhì)體后24小時(shí)，通過流式細(xì)胞術(shù)檢測外周血巨噬細(xì)胞數(shù)量。
清除結(jié)果：
圖1.腹腔注射給藥后巨噬細(xì)胞減少
參考文獻(xiàn)：Xiong X, Chen S, Shen J, et al. Cannabis suppresses antitumor immunity by inhibiting JAK/STAT signaling in T cells through CNR2. Signal Transduct Target Ther. 2022;7(1):99. Published 2022 Apr 6. doi:10.1038/s41392-022-00918-y(IF:18.187)
小鼠品系：雌性C57BL/6J小鼠
注射方式：腹腔注射
取樣部位：腹腔灌洗液
注射劑量：第0、3、6天分別給與15 mg/kg巨噬細(xì)胞清除劑
抗體選擇：CD11b，F(xiàn)4/80雙陽性
檢測方法：在第0、3、6天將氯膦酸脂質(zhì)體 (15 mg/kg)或懸浮在200 ml PBS中的空脂質(zhì)體腹腔注射到6周齡雌性C57BL/6J小鼠體內(nèi)，以消耗巨噬細(xì)胞，并在最后一次脂質(zhì)體注射后的第3、6天將100萬BMDMs懸浮在100 ml PBS中靜脈注射重建巨噬細(xì)胞，然后通過流式細(xì)胞術(shù)分析小鼠腹腔灌洗液巨噬細(xì)胞的損耗和重構(gòu)。
清除結(jié)果：
圖2.注射脂質(zhì)體后鼠腹腔灌洗液及巨噬細(xì)胞重建的流式分析
參考文獻(xiàn)：Zhang Z, Chen C, Yang F, et al. Itaconate is a lysosomal inducer that promotes antibacterial innate immunity [published online ahead of print, 2022 May 25. Mol Cell. 2022;S1097-2765(22)00443-9. doi:10.1016/j.molcel.2022.05.009 (IF:17.970)
小鼠品系：C57BL/6J
注射方式：靜脈注射
取樣部位：血液、乳腺
注射劑量：200 μl /只
抗體選擇：F4/80
清除方法：巨噬細(xì)胞耗竭模型采用氯膦酸脂質(zhì)體(0.15-0.2 mL/25 g)，給與200μL每只小鼠，靜脈注射于哺乳期小鼠體內(nèi)，消耗血液和乳腺內(nèi)巨噬細(xì)胞，在注射氯鈉脂質(zhì)體后24、48和72小時(shí)，通過流式細(xì)胞術(shù)檢測乳腺巨噬細(xì)胞數(shù)量。
清除結(jié)果：
圖3.a）血液學(xué)自動分析儀檢測血液中單核細(xì)胞(血液中巨噬細(xì)胞的一種形式)比例的變化；b）流式細(xì)胞術(shù)檢測乳腺巨噬細(xì)胞比例變化
參考文獻(xiàn)：Cai J, Peng J, Zang X, Feng J, Li R, Ren P, Zheng B, Wang J, Wang J, Yan M, Liu J, Deng R, Wang D. Mammary Leukocyte-Assisted Nanoparticle Transport Enhances Targeted Milk Trace Mineral Delivery. Adv Sci (Weinh). 2022 Sep;9(26):e2200841. doi: 10.1002/advs.202200841. Epub 2022 Jun 30. PMID: 35773238; PMCID: PMC9475556. (IF:17.521)
小鼠品系：C57BL/6
注射方式：腹腔注射
取樣部位：外周血
注射劑量：200 μl /只
抗體選擇：F4/80
檢測方法： 第3天腹腔注射氯磷酸脂質(zhì)體200 μL，去除巨噬細(xì)胞。用流式細(xì)胞術(shù)檢測巨噬細(xì)胞缺失效應(yīng)。
清除結(jié)果：
圖4.免疫細(xì)胞缺失流式分析
參考文獻(xiàn)：Yu X, Long Y, Chen B, et al. PD-L1/TLR7 dual-targeting nanobody-drug conjugate mediates potent tumor regression via elevating tumor immunogenicity in a host-expressed PD-L1 bias-dependent way. J Immunother Cancer. 2022;10(10):e004590. doi:10.1136/jitc-2022-004590(IF:12.469)
產(chǎn)品簡介
產(chǎn)品組成：1 mL脂質(zhì)體體懸液（1.5-2μm）中所含的氯膦酸大約為5 mg，所用的磷酸鹽緩沖液為10 mM Na2HPO4，10 mM NaH2PO4，140 mM NaCl。
產(chǎn)品列表
部分參考文獻(xiàn)列表
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