Waller E K, Olweus J, Lund-Johansen F, Huang S, Nguyen M, Guo G R, Terstappen L
Becton Dickinson Immunocytometry Systems, San Jose, CA, USA.
Blood. 1995 May 1;85(9):2422-35.
There is a long-standing controversy as to whether a single bone marrow (BM)-derived cell can differentiate along both hematopoietic and stromal lineages. Both primitive hematopoietic and stromal progenitor cells in human BM express the CD34 antigen but lack expression of other surface markers, such as CD38. In this study we examined the CD34+, CD38- fraction of human fetal BM by multiparameter fluorescence-activated cell sorting (FACS) analysis and single-cell sorting. CD34+, C38- cells could be divided into HLA-DR+ and HLA-DR- fractions. After single-cell sorting, 59% of the HLA-DR+ cells formed hematopoietic colonies. In contrast, the CD34+, CD38-, HLA-DR- cells were much more heterogeneous with respect to their light scatter properties, expression of other hematopoietic markers (CD10, CD36, CD43, CD49b, CD49d, CD49e, CD50, CD62E, CD90w, CD105, and CD106), and growth properties. Single CD34+, CD38-, HLA-DR- cells sorted into individual culture wells formed either hematopoietic or stromal colonies. The presence or absence of CD50 (ICAM-3) expression distinguished hematopoietic from stromal progenitors within the CD34+, CD38-, HLA-DR- population. The CD50+ fraction had light scatter characteristics and growth properties of hematopoietic progenitor cells. In contrast, the CD50- fraction lacked hematopoietic progenitor activity but contained clonogenic stromal progenitors at a mean frequency of 5%. We tested the hypothesis that cultures derived from single cells with the CD34+, CD38-, HLA-DR- phenotype could differentiate along both a hematopoietic and stromal lineage. The cultures contained a variety of mesenchymal cell types and mononuclear cells that had the morphologic appearance of histiocytes. Immunophenotyping of cells from these cultures indicated a stromal rather than a hematopoietic origin. In addition, the growth of the histiocytic cells was independent of the presence or the absence of hematopoietic growth factors. Based on sorting more than 30,000 single cells with the CD34+, CD38-, HLA-DR- phenotype into individual culture wells, and an analysis of 864 stromal cultures initiated by single CD34+ BM cells, this study does not support the hypothesis of a single common progenitor for both hematopoietic and stromal lineages within human fetal BM.
关于单个骨髓（BM）来源的细胞是否能同时沿着造血和基质谱系分化，一直存在争议。人类BM中的原始造血祖细胞和基质祖细胞均表达CD34抗原，但缺乏其他表面标志物的表达，如CD38。在本研究中，我们通过多参数荧光激活细胞分选（FACS）分析和单细胞分选，对人类胎儿BM的CD34 +、CD38 - 部分进行了检测。CD34 +、C38 - 细胞可分为HLA - DR + 和HLA - DR - 部分。单细胞分选后，59%的HLA - DR + 细胞形成造血集落。相比之下，CD34 +、CD38 -、HLA - DR - 细胞在光散射特性、其他造血标志物（CD10、CD36、CD43、CD49b、CD49d、CD49e、CD50、CD62E、CD90w、CD105和CD106）的表达以及生长特性方面更为异质。分选到单个培养孔中的单个CD34 +、CD38 -、HLA - DR - 细胞形成造血或基质集落。CD50（ICAM - 3）表达的有无区分了CD34 +、CD38 -、HLA - DR - 群体中的造血祖细胞和基质祖细胞。CD50 + 部分具有造血祖细胞的光散射特征和生长特性。相比之下，CD50 - 部分缺乏造血祖细胞活性，但含有克隆形成性基质祖细胞，平均频率为5%。我们检验了这样一个假设，即源自具有CD34 +、CD38 -、HLA - DR - 表型的单细胞的培养物可以沿着造血和基质谱系分化。这些培养物包含多种间充质细胞类型和具有组织细胞形态外观的单核细胞。对这些培养物中的细胞进行免疫表型分析表明其起源于基质而非造血。此外，组织细胞的生长与造血生长因子的有无无关。基于将超过30000个具有CD34 +、CD38 -、HLA - DR - 表型的单细胞分选到单个培养孔中，并对由单个CD34 + BM细胞启动的864个基质培养物进行分析，本研究不支持人类胎儿BM中造血和基质谱系存在单一共同祖细胞的假设。
