Comparison of the characteristics of macrophages derived from murine spleen, peritoneal cavity, and bone marrow

Abstract

Macrophages have a diverse set of functions based upon their activation states. The activation states, including resting (M0) and polarizing (M1 and M2) states, of macrophages derived from the mouse bone marrow, spleen, and peritoneal cavity (BMs, SPMs, and PCMs, respectively) were compared. We evaluated the macrophage yield per mouse and compared the surface markers major histocompatibility complex (MHC) II and CD86 by flow cytometry. The relative mRNA levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, mannose receptor (MR), and Ym1 in the M0, M1, and M2 states were also compared using real-time polymerase chain reaction (PCR) analysis. Bone marrow yielded the most macrophages with the best homogeneity, but they were polarized toward the M2 phenotype. All three types of macrophages had the capacity to polarize into the M1 and M2 states, but SPMs had a stronger capacity to polarize into M1. The three types of macrophages showed no differences in their capacity to polarize into the M2 state. Therefore, the three types of macrophages have distinct characteristics regardless of their resting or polarizing states. Although bone marrow can get large amounts of homogeneous macrophages, the macrophages cannot replace tissue-derived macrophages.

中文概要

目的

对比分析脾脏、腹腔和骨髓源性巨噬细胞(SPMs、PCMs 和BMs)在安静及极化状态下的差异。

创新点

首次对比分析脾脏、腹腔和骨髓源性巨噬细胞在安静(M0)及极化状态(M1 和M2)下的特征。

方法

通过小鼠脾脏研磨及单细胞贴壁获得脾源性巨噬细胞;腹腔灌洗及细胞贴壁获得腹腔源性巨噬细胞;骨髓贴壁细胞在巨噬细胞集落刺激因子体外刺激下培养7 天获得骨髓源性巨噬细胞。三种细胞即为M0 型巨噬细胞,M0 在干扰素及脂多糖刺激下获得M1 型巨噬细胞,M0 在白介素4(IL-4)刺激下获得M2 型巨噬细胞。通过流式细胞仪分析三种类型巨噬细胞在三种状态下的II 类主要组织相容性复合体(MHC II)和CD86 表达差异。通过实时荧光定量聚合酶链式反应(qPCR)检测肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、甘露糖受体(MR)和类几丁质酶3 样分子(Ym1)的表达变化。

结论

骨髓贴壁细胞培养能获得最大量的同源巨噬细胞(图1 和2),但表型偏向于M2 型巨噬细胞(图3 和4)。三种巨噬细胞均能极化为M1 和M2 型巨噬细胞(图5),其中SPMs 具有更强的M1 型极化能力,而M2 型极化能力之间未见明显差异(图6)。综上所述,三种细胞无论在安静及极化状态下均具有不同的特征,骨髓可以获得大量同源性巨噬细胞,但性质不同于组织源性的脾脏和腹腔巨噬细胞。