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基于聚糖的细胞特征分析和分离方法：以造血为例。

A glycan-based approach to cell characterization and isolation: Hematopoiesis as a paradigm.

机构信息

Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY.

Department of Genetics, Albert Einstein College of Medicine, Bronx, NY.

出版信息

J Exp Med. 2022 Nov 7;219(11). doi: 10.1084/jem.20212552. Epub 2022 Sep 6.

DOI:10.1084/jem.20212552
PMID:36066492
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9455685/
Abstract

Cell surfaces display a wide array of molecules that confer identity. While flow cytometry and cluster of differentiation (CD) markers have revolutionized cell characterization and purification, functionally heterogeneous cellular subtypes remain unresolvable by the CD marker system alone. Using hematopoietic lineages as a paradigm, we leverage the extraordinary molecular diversity of heparan sulfate (HS) glycans to establish cellular "glycotypes" by utilizing a panel of anti-HS single-chain variable fragment antibodies (scFvs). Prospective sorting with anti-HS scFvs identifies functionally distinct glycotypes within heterogeneous pools of mouse and human hematopoietic progenitor cells and enables further stratification of immunophenotypically pure megakaryocyte-erythrocyte progenitors. This stratification correlates with expression of a heptad of HS-related genes that is reflective of the HS epitope recognized by specific anti-HS scFvs. While we show that HS glycotyping provides an orthogonal set of tools for resolution of hematopoietic lineages, we anticipate broad utility of this approach in defining and isolating novel, viable cell types across diverse tissues and species.

摘要

细胞表面展示出广泛的分子，赋予其身份。虽然流式细胞术和分化群（CD）标志物已经彻底改变了细胞特征和纯化，但仅凭 CD 标志物系统，功能上异质的细胞亚群仍然无法解决。我们利用肝素硫酸（HS）聚糖的非凡分子多样性，利用一组抗 HS 单链可变片段抗体（scFv），将其作为范例，建立细胞“糖型”。利用抗 HS scFv 进行前瞻性分选，可以在异质的小鼠和人类造血祖细胞群体中识别出功能不同的糖型，并能够进一步对免疫表型纯巨核细胞-红细胞祖细胞进行分层。这种分层与 HS 相关基因的七个基因的表达相关，这些基因反映了特定抗 HS scFv 识别的 HS 表位。虽然我们表明 HS 糖型为解析造血谱系提供了一组正交工具，但我们预计这种方法在定义和分离不同组织和物种中的新型、可行细胞类型方面具有广泛的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/a2a5ce9d3d7a/JEM_20212552_Fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/81d019b815d5/JEM_20212552_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/6770b5a319a8/JEM_20212552_FigS1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/0f10f5fd55a9/JEM_20212552_FigS2.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/cf16827e5eab/JEM_20212552_FigS3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/29c34b4a8186/JEM_20212552_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/5635cf0e866b/JEM_20212552_Fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/01df270e0dd3/JEM_20212552_Fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/75d3d9986906/JEM_20212552_FigS4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/20847ce3617e/JEM_20212552_FigS5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/aa5478ac4a79/JEM_20212552_Fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/a2a5ce9d3d7a/JEM_20212552_Fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/81d019b815d5/JEM_20212552_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/6770b5a319a8/JEM_20212552_FigS1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/0f10f5fd55a9/JEM_20212552_FigS2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/18d704c75671/JEM_20212552_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/2f7054eda3a3/JEM_20212552_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/cf16827e5eab/JEM_20212552_FigS3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/29c34b4a8186/JEM_20212552_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/5635cf0e866b/JEM_20212552_Fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/01df270e0dd3/JEM_20212552_Fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/75d3d9986906/JEM_20212552_FigS4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/20847ce3617e/JEM_20212552_FigS5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/aa5478ac4a79/JEM_20212552_Fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0114/9455685/a2a5ce9d3d7a/JEM_20212552_Fig8.jpg

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