肝臟巨噬細(xì)胞和骨髓來源巨噬細(xì)胞流式區(qū)分圈門策略

為了研究腫瘤細(xì)胞早期定植。中期生長和晚期轉(zhuǎn)移等階段以及肝臟定居巨噬細(xì)胞KF和骨髓來源巨噬細(xì)胞BMDM等巨噬細(xì)胞的功能作用。研究者使用了門靜脈注射胰腺導(dǎo)管腺癌PDACYFP-cells模型。三個(gè)階段，兩種巨噬細(xì)胞。如何區(qū)分肝臟定居巨噬細(xì)胞KF和骨髓來源巨噬細(xì)胞BMDM？研究者在模型里面找到差異表達(dá)基因clec4f。

CLEC4F; CLECSF13; C-type (calcium dependent, carbohydrate-recognition domain) lectin, superfamily member 13; C-type lectin 13; C-type lectin domain family 4, member F; Galactose Particle Receptor; KCLR; KCR。CLEC4F/CLECSF13 蛋白是一種半乳糖和巖藻糖結(jié)合受體，表明其在內(nèi)吞作用中發(fā)揮作用，表明它可能參與內(nèi)化各種細(xì)胞成分并促進(jìn)重要的細(xì)胞過程。

CLEC4F (C-type lectin domain; family 4, member F; also known as the Kupffer cell receptor and fucose receptor) is an 80 kDa, type II transmembrane glycoprotein member of the C-type lectin superfamily. Mature mouse CLEC4F consists of a 42 amino acid (aa) cytoplasmic domain, a 27 aa transmembrane segment, and a 479 aa extracellular domain (ECD) that contains an extended stalk region plus one carbohydrate recognition domain . Within the ECD, mouse CLEC4F shares 48% and 79% aa sequence identity with human and rat CLEC4F, respectively. The stalk region of CLEC4F is a coiled coil domain that mediates homotrimer formation. CLEC4F is expressed on Kupffer cells in the liver, but not on macrophages in other tissues. CLEC4F preferentially binds galactose and N?acetylgalactosamine in a calcium-dependent manner. Its activity at neutral, but not at acidic pH, suggests a capacity to internalize and release ligands into the endosomal system.

如何通過流式區(qū)分：肝臟巨噬細(xì)胞和骨髓來源巨噬細(xì)胞流式區(qū)分圈門策略見下圖。如下是圈門策略。

Flow Cytometry gating schema. a, Representative dot plots showing gating strategy for identifying macrophage populations in the liver. Gates were based on FMOs. Macrophages were defined as live, single cells, expressing CD45, CD11b, and F4/80 and lacking expression of CD3, CD19, CD11c, and Ly6G. Kupffer cells (KC) were distinguished from bone marrow derived macrophages (BMDM) based on Clec4f expression. Staining is representative of a liver sample from a C57BL/6 mouse. b, Representative FCM gating strategies for detection of YFP+ tumor cells in the liver after iPo injection of PDACYFP cells. Populations shown were gated on single, live cells.

Clodronate Liposomes氯膦酸鹽脂質(zhì)體清除肝臟巨噬細(xì)胞，疾病模型為：門靜脈注射PDACYFP-cells。PDAC是胰腺導(dǎo)管腺癌（Pancreatic Ductal Adenocarcinoma）的縮寫，屬于胰腺癌中常見且惡性程度高的類型，占胰腺惡性腫瘤的85%-90%。它起源于胰腺導(dǎo)管上皮細(xì)胞，侵襲性強(qiáng)、進(jìn)展迅速，多數(shù)患者確診時(shí)已處于中晚期，5年生存率不足10%。荷蘭Liposoma巨噬細(xì)胞清除劑Clodronate Liposomes見刊于Nature Communications：Kupffer 細(xì)胞可防止小鼠胰腺導(dǎo)管腺癌轉(zhuǎn)移到肝臟。Clec4f可以區(qū)分肝臟巨噬細(xì)胞KF和骨髓來源巨噬細(xì)胞BMDM嗎？詳細(xì)類容可以見如下文獻(xiàn)。

論文信息：

論文題目：Kupffer cells prevent pancreatic ductal adenocarcinoma metastasis to the liver in mice

期刊名稱：Nature Communications

時(shí)間期卷：14, Article number: 6330 (2023)

在線時(shí)間：2023年10月10日

DOI：doi.org/10.1038/s41467-023-41771-z

產(chǎn)品信息：

貨號(hào)：C-010

規(guī)格：10ml

品牌：Liposoma

產(chǎn)地：荷蘭

名稱：Clodronate Liposomes

辦事處：Target Technology(靶點(diǎn)科技)