Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time.
Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time.
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DOI：
10.3389/fimmu.2022.1064084
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发表时间：
2022
影响因子：
7.3
通讯作者：
中科院分区：
文献类型：
--
作者：
以鸡骨髓来源的巨噬细胞(Φ)和树突状细胞(BMDC)为模型，对单核吞噬系统(MPS)进行了研究。一种广泛使用的体外培养巨噬细胞和DC的方法是在集落刺激因子1的存在下培养骨髓细胞来分化骨髓间充质干细胞，并用粒细胞巨噬细胞集落刺激因子2和白介素4来分化骨髓间充质干细胞，而单用集落刺激因子2可以诱导粒细胞巨噬细胞衍生DC的形成。然而，在鸡中，这些培养物所代表的MPS细胞谱系及其功能还知之甚少。在这里，我们破译了鸡骨髓基质细胞Φ和骨髓基质细胞树突状细胞在表型、功能和转录方面的差异，并检测了在培养第2、4、6和8天在没有IL-4的情况下生长的DC细胞的差异。骨髓间充质干细胞和骨髓间充质干细胞培养产生形态上不同的细胞培养物，而骨髓间充质干细胞和骨髓间充质干细胞培养物则发育成形态上均一的细胞群体。在表型水平上，所有培养的MHCII、CD11c和CSF1R转基因的细胞百分比和表达水平相似，而在骨髓基质细胞Φ中，MRC1L-B的表达随着时间的推移而降低。所有的培养物都能有效地摄取0.5微米的珠子，但吞噬1微米的珠子的能力很差。在分析的每一天，不同培养物的吞噬体酸化动力学几乎没有差异。时间转录分析表明，所有培养物都表达与哺乳动物巨噬细胞相关的CSF3R、MERTK、SEPP1、SPI1和TLR4基因。相反，在BMDC和GMDC中，与DC相关的基因Flt3、XCR1和CAMD1在第2天低水平表达，之后表达水平下降。总的来说，鸡CSF2+IL-4-和CSF2依赖的BM培养代表巨噬细胞谱系的细胞，而不是诱导传统的DC。
Chicken bone marrow-derived macrophages (BMMΦ) and dendritic cells (BMDC) are utilized as models to study the mononuclear phagocytic system (MPS). A widely used method to generate macrophages and DC in vitro is to culture bone marrow cells in the presence of colony-stimulating factor-1 (CSF1) to differentiate BMMΦ and granulocyte-macrophage-CSF (GM-CSF, CSF2) and interleukin-4 (IL-4) to differentiate BMDC, while CSF2 alone can lead to the development of granulocyte-macrophage-CSF-derived DC (GMDC). However, in chickens, the MPS cell lineages and their functions represented by these cultures are poorly understood. Here, we decipher the phenotypical, functional and transcriptional differences between chicken BMMΦ and BMDC along with examining differences in DC cultures grown in the absence of IL-4 on days 2, 4, 6 and 8 of culture. BMMΦ cultures develop into a morphologically homogenous cell population in contrast to the BMDC and GMDC cultures, which produce morphologically heterogeneous cell cultures. At a phenotypical level, all cultures contained similar cell percentages and expression levels of MHCII, CD11c and CSF1R-transgene, whilst MRC1L-B expression decreased over time in BMMΦ. All cultures were efficiently able to uptake 0.5 µm beads, but poorly phagocytosed 1 µm beads. Little difference was observed in the kinetics of phagosomal acidification across the cultures on each day of analysis. Temporal transcriptomic analysis indicated that all cultures expressed high levels of CSF3R, MERTK, SEPP1, SPI1 and TLR4, genes associated with macrophages in mammals. In contrast, low levels of FLT3, XCR1 and CAMD1, genes associated with DC, were expressed at day 2 in BMDC and GMDC after which expression levels decreased. Collectively, chicken CSF2 + IL-4- and CSF2-dependent BM cultures represent cells of the macrophage lineage rather than inducing conventional DC.
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影响因子：
2.2
作者：
Foulon, Eliane;Foucras, Gilles
通讯作者：
Foucras, Gilles
影响因子：
4.8
作者：
Dhami, R;Schuchman, EH
通讯作者：
Schuchman, EH
影响因子：
30.5
作者：
通讯作者：
--
影响因子：
30.5
作者：
Erlich, Ziv;Shlomovitz, Inbar;Gerlic, Motti
通讯作者：
Gerlic, Motti
影响因子：
15.3
作者：
Celada, A;Borras, FE;Maki, RA
通讯作者：
Maki, RA