National Heart, Lung and Blood Institute, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892, USA.
Blood. 2011 Feb 3;117(5):1550-4. doi: 10.1182/blood-2009-03-212803. Epub 2010 Dec 16.
Various combinations of antibodies directed to cell surface markers have been used to isolate human and rhesus macaque hematopoietic stem cells (HSCs). These protocols result in poor enrichment or require multiple complex steps. Recently, a simple phenotype for HSCs based on cell surface markers from the signaling lymphocyte activation molecule (SLAM) family of receptors has been reported in the mouse. We examined the possibility of using the SLAM markers to facilitate the isolation of highly enriched populations of HSCs in humans and rhesus macaques. We isolated SLAM (CD150(+)CD48(-)) and non-SLAM (not CD150(+)CD48(-)) cells from human umbilical cord blood CD34(+) cells as well as from human and rhesus macaque mobilized peripheral blood CD34(+) cells and compared their ability to form colonies in vitro and reconstitute immune-deficient (nonobese diabetic/severe combined immunodeficiency/interleukin-2 γc receptor(null), NSG) mice. We found that the CD34(+) SLAM population contributed equally or less to colony formation in vitro and to long-term reconstitution in NSG mice compared with the CD34(+) non-SLAM population. Thus, SLAM family markers do not permit the same degree of HSC enrichment in humans and rhesus macaques as in mice.
各种针对细胞表面标记物的抗体组合已被用于分离人和恒河猴造血干细胞(HSCs)。这些方案导致富集效果不佳或需要多个复杂步骤。最近,在小鼠中报道了一种基于信号淋巴细胞激活分子(SLAM)受体家族的细胞表面标记物的简单 HSC 表型。我们研究了使用 SLAM 标记物来促进人源和恒河猴来源的高纯度 HSC 群体分离的可能性。我们从人脐带血 CD34+细胞以及人源和恒河猴动员外周血 CD34+细胞中分离出 SLAM(CD150+CD48-)和非-SLAM(非 CD150+CD48-)细胞,并比较它们在体外形成集落和重建免疫缺陷(非肥胖糖尿病/严重联合免疫缺陷/白细胞介素-2γc 受体(null),NSG)小鼠的能力。我们发现,与 CD34+非-SLAM 群体相比,CD34+ SLAM 群体在体外形成集落和在 NSG 小鼠中进行长期重建的能力相等或更低。因此,SLAM 家族标记物不能像在小鼠中那样在人类和恒河猴中实现相同程度的 HSC 富集。