Charbord Pierre, Oostendorp Robert, Pang Wenxin, Hérault Olivier, Noel Frederic, Tsuji Takashi, Dzierzak Elaine, Péault Bruno
Inserm U506, Hopital Paul Brousse, Villejuif, France.
Exp Hematol. 2002 Oct;30(10):1202-10. doi: 10.1016/s0301-472x(02)00895-0.
To better understand the differentiation of stromal cells of the hematopoietic microenvironment, we set out to characterize stromal cells from the different developmental sites of hematopoiesis in the mouse (30 bone marrow, 7 spleen, 3 embryonic and 15 fetal liver, 6 yolk sac, and 6 aorta-gonad-mesonephros lines) for expression of 22 cytoskeletal, membrane, and extracellular matrix proteins.
Western blotting, immunofluorescence, and flow cytometry were used. Statistical methods included principal components analysis and analysis of variance.
Stromal cells from 11 dpc mouse embryos express mesenchymal and vascular smooth muscle cell (VSMC) markers. Principal components analysis on the 70 stromal cell lines isolated from different anatomic sites and developmental stages allows classification of stromal lines along a mesenchymal to VSMC differentiation pathway. Stromal cells do not express endothelial and hematopoietic differentiation membrane antigens, but they do express integrin alpha(5), alpha(6), and beta(1) subunits, vascular cell adhesion molecule-1, CD44, stem cell antigen-1, Thy-1, CD34, and endoglin. The intensity of expression of certain markers differs between lines according to the anatomic site of origin.
This study indicates that stromal cells, whatever their anatomic site of origin, follow a VSMC differentiation pathway, suggesting a blood-forming tissue-specific differentiation of mesenchymal stem cells. Differential quantitative expression of distinct sets of markers appears to be correlated with the anatomic sites of origin of the stromal cells.
为了更好地理解造血微环境中基质细胞的分化，我们着手对来自小鼠造血不同发育部位（30个骨髓、7个脾脏、3个胚胎、15个胎肝、6个卵黄囊和6个主动脉-性腺-中肾系）的基质细胞进行表征，以检测22种细胞骨架、膜和细胞外基质蛋白的表达情况。
采用蛋白质免疫印迹法、免疫荧光法和流式细胞术。统计方法包括主成分分析和方差分析。
妊娠第11天小鼠胚胎的基质细胞表达间充质和血管平滑肌细胞（VSMC）标志物。对从不同解剖部位和发育阶段分离出的70个基质细胞系进行主成分分析，可将基质细胞系沿着间充质到VSMC的分化途径进行分类。基质细胞不表达内皮和造血分化膜抗原，但它们表达整合素α(5)、α(6)和β(1)亚基、血管细胞黏附分子-1、CD44、干细胞抗原-1、Thy-1、CD34和内皮糖蛋白。某些标志物的表达强度在不同细胞系之间因起源的解剖部位而异。
本研究表明，无论起源于何处，基质细胞都遵循VSMC分化途径，提示间充质干细胞存在造血组织特异性分化。不同标志物组的差异定量表达似乎与基质细胞的起源解剖部位相关。
