Hinchly Taylor, Bonnet Dominique, Anjos-Afonso Fernando
Haematopoietic Signalling Group, European Cancer Stem Cell Institute, School of Biosciences, Cardiff University, Cardiff, United Kingdom.
Haematopoietic Stem Cell Lab, The Francis Crick Institute, London, United Kingdom.
Exp Hematol. 2025 Apr;144:104729. doi: 10.1016/j.exphem.2025.104729. Epub 2025 Jan 30.
Recently, human CD34 hematopoietic stem cells (HSCs) have been purified to a frequency of approximately one in three cells, a population denoted as CD34CD38CD45RACD90 endothelial protein C receptor (EPCR) HSCs. This work aimed to evaluate the methodology for CD34 HSC isolation, exploring differences in antibody clones, conjugates, source of cells, and additional cell surface antigens (integrin-α6, CLEC9A, and GPRC5C) to enhance the purity of these EPCR HSCs. We are emphasizing here the importance of experimental planning and antibody panel selection concerning the isolation of these human HSCs from multiple sources and providing important notes on the pitfalls of the reagents used for such purposes. Our results should enable a better reproducibility of results between laboratory tests as well as further pursuits of work toward improving the enrichment of human HSCs.
最近，人类CD34造血干细胞（HSCs）已被纯化至大约每三个细胞中有一个的频率，该群体被称为CD34⁺CD38⁻CD45RA⁻CD90⁺内皮蛋白C受体（EPCR）⁺ HSCs。这项工作旨在评估CD34⁺ HSC分离方法，探索抗体克隆、偶联物、细胞来源以及其他细胞表面抗原（整合素-α6、CLEC9A和GPRC5C）的差异，以提高这些EPCR⁺ HSCs的纯度。在此，我们强调了实验规划和抗体组合选择对于从多种来源分离这些人类HSCs的重要性，并提供了有关用于此类目的试剂陷阱的重要注意事项。我们的结果应能使实验室测试之间的结果具有更好的可重复性，并有助于进一步开展工作以改善人类HSCs的富集。
