A simple method to increase the proportion of bonemarrow-derived macrophages positive for M-CSFR using the reducing agent dithiothreitol (DTT)
A simple method to increase the proportion of bonemarrow-derived macrophages positive for M-CSFR using the reducing agent dithiothreitol (DTT)
复制标题
使用还原剂二硫苏糖醇 (DTT) 增加骨髓源性巨噬细胞 M-CSFR 阳性比例的简单方法
DOI：
10.1016/j.mex.2018.11.014
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Katoh-Y
中科院分区：
文献类型：
--
作者：
Hashimoto-R;Daida-H;Okada-T;Katoh-Y
只有少数骨髓源性巨噬细胞（BM-MF）对巨噬细胞集落刺激因子受体（M-CSFR）呈阳性。因此，需要一种方法来增加对M-CSFR呈阳性的BM-MF的比例，以便于研究M-CSFR下调对各种疾病的影响。我们使用小鼠原代BM-MF，使用流式细胞术评估细胞质膜上M-CSFR的表达。用还原剂二硫苏糖醇（DTT）处理，增加了M-CSFR阳性的BM-MF的比例，并且这种增加是时间依赖性的。我们接下来确定DTT处理的BM-MF是否可以导致M-CSFR的下调。用脂多糖（LPS）处理24？H.降低DTT处理的BM-MF中M-CSFR阳性的比例。这些结果表明，DTT处理增加了对M-CSFR呈阳性的BM-MF的比例，并且用LPS处理可以消除M-CSFR对BM-MF的上调。在这里，我们提出了一种使用还原剂DTT增加M-CSFR阳性BM-MF数量的简单方法，这可能有助于研究M-CSFR下调与某些疾病之间的关系。DTT处理后，在膜上表达M-CSFR的BM-MF的比例增加约两倍。
Only a few bone marrow-derived macrophages (BM-MF) are positive for macrophage colony-stimulating factor receptor (M-CSFR). Thus, a method is needed to increase the proportion of BM-MF that are positive for M-CSFR to facilitate the investigation of the effects of M-CSFR downregulation on various diseases. We used mouse primary BM-MF to evaluate the expression of M-CSFR on the cytoplasmic membrane using flow cytometry. Treatment with a reducing agent, dithiothreitol (DTT), increased the proportion of BM-MF that were positive for M-CSFR, and this increase was time dependent. We next determined whether DTT-treated BM-MF can lead to the downregulation of M-CSFR. Treatment with lipopolysaccharide (LPS) for 24?h. decreased the proportion of DTT-treated BM-MF that were positive for M-CSFR. These results suggest that DTT treatment increases the proportion of BM-MF that are positive for M-CSFR and that the upregulation of M-CSFR on BM-MF can be abrogated by treatment with LPS. Here, we propose a simple method to increase the number of M-CSFR-positive BM-MF using the reducing agent DTT, which could be useful in investigations of the relationship between the downregulation of M-CSFR and some diseases.•The proportion of BM-MF that expresses M-CSFR on the membrane increases by approximately twice following DTT treatment.