Generation of eosinophils from unselected bone marrow progenitors: wild-type, TLR- and eosinophil-deficient mice.

Kimberly D Dyer, Caroline M Percopo, Helene F Rosenberg
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引用次数: 11

Abstract

We have recently devised a culture method that generates large numbers of eosinophils at high purity from unselected BALB/c mouse bone marrow progenitors [Dyer et al., 2008. J. Immunol. 181: 4004-9]. Here we present the extended scope of this approach, as we have used this method successfully to generate eosinophil cultures of virtually 100% purity from bone marrow from C57BL/6 mice, and from TLR2, TLR3, TLR7 and TLR9-gene-deleted mouse strains on the C57BL/6 background. Both wild-type and TLR3 gene-deleted bone marrow eosinophils (bmEos) are functional, releasing peroxidase in response to the secretogogue, platelet activating factor. We have also used this method to re-evaluate production of eosinophils in bone marrow cultures from ΔdblGATA mice, a strain that is eosinophil-deficient in vivo. Interestingly, bmEos can be detected in the ΔdblGATA cultures (5% of total cells at day 10), although ~80-fold fewer bmEos are detected in ΔdblGATA than in parallel wild-type (BALB/c) bone marrow cultures. Overall, we find that generation of large numbers of eosinophils at high purity from unselected bone marrow progenitors proceeds efficiently in a variety of wild-type and gene-deleted strains, and as such this approach shows promise as a universal method for the study of eosinophil structure and function.

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从未选择的骨髓祖细胞产生嗜酸性粒细胞:野生型,TLR-和嗜酸性粒细胞缺陷小鼠。
我们最近设计了一种培养方法,可以从未选择的BALB/c小鼠骨髓祖细胞中产生大量高纯度的嗜酸性粒细胞[Dyer等人,2008]。[j].中国生物医学工程学报,2016,33(2):444 - 444。在这里,我们展示了这种方法的扩展范围,因为我们已经使用这种方法成功地从C57BL/6小鼠的骨髓中,以及在C57BL/6背景下的TLR2, TLR3, TLR7和tlr9基因缺失的小鼠株中产生几乎100%纯度的嗜酸性粒细胞培养物。野生型和TLR3基因缺失的骨髓嗜酸性粒细胞(bmEos)都是功能性的,在分泌剂和血小板激活因子的作用下释放过氧化物酶。我们也用这种方法重新评估了ΔdblGATA小鼠骨髓培养中嗜酸性粒细胞的产生,这是一种体内嗜酸性粒细胞缺乏的菌株。有趣的是,在ΔdblGATA培养物中可以检测到bmEos(第10天占总细胞的5%),尽管ΔdblGATA中检测到的bmEos比平行野生型(BALB/c)骨髓培养物少约80倍。总的来说,我们发现在各种野生型和基因缺失菌株中,从未选择的骨髓祖细胞中产生大量高纯度的嗜酸性粒细胞是有效的,因此这种方法有望成为研究嗜酸性粒细胞结构和功能的通用方法。
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