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Updated: May 26, 2025

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从全血中分离和描述低密度和正常密度中性粒细胞

Anjali S Yennemadi¹, Joseph Keane¹, Gina Leisching²

¹TB Immunology Group, Department of Clinical Medicine, Trinity Translational Medicine Institute, St James's Hospital, Trinity College Dublin, The University of Dublin.

Journal of visualized experiments : JoVE

|February 25, 2025

概括

研究人员开发了一种新方法,从人类血液中分离低密度中性粒细胞 (LDNs) 和正常密度中性粒细胞 (NDNs). 这种技术改善了研究中性粒细胞在健康和疾病中的异质性.

科学领域:

免疫学免疫学免疫学
血液学血液学血液学
细胞生物学细胞生物学

背景情况:

循环中性粒细胞是多样化的,在历史上被研究为一个单一的群体.
低密度中性粒细胞 (LDNs) 和正常密度中性粒细胞 (NDNs) 呈现出不同的功能和代谢特征.
准确地分离中性粒细胞亚型对于理解它们在生理学和病理学中的作用至关重要.

研究的目的:

提出一种经过修改,未经改进的方法,用于从人体全血中分离和分离LDN和NDN.
为了使不同中性粒细胞亚群的下游分析更加精确.
促进对中性粒细胞异质性及其影响的理解.

主要方法:

用酸盐缓冲盐水 (PBS) 制备了一种密度梯度介质 (1.135 g/mL),以创建55%,70%和81%的密度梯度.
从外围血液中用负选择磁性套件分离出总中性粒细胞,并在55%的分数中重新悬浮.
密度梯度被离心,在特定的接口产生明显的LDN和NDN波段,随后通过流细胞计 (CD14lo CD15+SSChi) 进行细胞采集,洗和纯度评估.

主要成果:

建立了一种可靠的方法来将LDN和NDN与外周血液分离.
对于LDN和NDN的分离纯度被确定为≥93%的活细胞.
该方法确保了孤立的中性粒细胞亚群的高纯度和活力.

结论:

这种方法提供了一种有效的方法,可以对LDN和NDN进行完整的隔离和分离.
提高中性粒细胞分离的精度支持更准确的下游分析.
这种技术对于在各种环境中推进对中性粒细胞异质性的研究至关重要.

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Overview Of Cell Separation And Isolation

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Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.