Martinovic Snjezana, Mazic Sanja, Kisic Veronika, Basic Nikolina, Jakic-Razumovic Jasminka, Borovecki Fran, Batinic Drago, Simic Petra, Grgurevic Lovorka, Labar Boris, Vukicevic Slobodan
Department of Anatomy, Medical School University of Zagreb, Salata 11, Croatia.
J Histochem Cytochem. 2004 Sep;52(9):1159-67. doi: 10.1369/jhc.4A6263.2004.
Highly purified primitive hemopoietic stem cells express BMP receptors but do not synthesize bone morphogenetic proteins (BMPs). However, exogenously added BMPs regulate their proliferation, differentiation, and survival. To further explore the mechanism by which BMPs might be involved in hemopoietic differentiation, we tested whether stromal cells from long-term culture (LTC) of normal human bone marrow produce BMPs, BMP receptors, and SMAD signaling molecules. Stromal cells were immunohistochemically characterized by the presence of lyzozyme, CD 31, factor VIII, CD 68, S100, alkaline phosphatase, and vimentin. Gene expression was analyzed by RT-PCR and the presence of BMP protein was confirmed by immunohistochemistry (IHC). The supportive role of the stromal cell layer in hemopoiesis in vitro was confirmed by a colony assay of clonogenic progenitors. Bone marrow stromal cells express mRNA and protein for BMP-3, -4, and -7 but not for BMP-2, -5, and -6 from the first to the eighth week of culture. Furthermore, stromal cells express the BMP type I receptors, activin-like kinase-3 (ALK-3), ALK-6, and the downstream transducers SMAD-1, -4, and -5. Thus, human bone marrow stromal cells synthesize BMPs, which might exert their effects on hemopoietic stem cells in a paracrine manner through specific BMP receptors.
高度纯化的原始造血干细胞表达骨形态发生蛋白（BMP）受体，但不合成骨形态发生蛋白（BMP）。然而，外源性添加的BMP可调节其增殖、分化和存活。为了进一步探索BMP可能参与造血分化的机制，我们检测了正常人骨髓长期培养（LTC）的基质细胞是否产生BMP、BMP受体和SMAD信号分子。通过溶菌酶、CD 31、因子VIII、CD 68、S100、碱性磷酸酶和波形蛋白的存在对基质细胞进行免疫组织化学鉴定。通过逆转录聚合酶链反应（RT-PCR）分析基因表达，并通过免疫组织化学（IHC）确认BMP蛋白的存在。通过克隆祖细胞集落测定证实了基质细胞层在体外造血中的支持作用。在培养的第一至第八周，骨髓基质细胞表达BMP-3、-4和-7的mRNA和蛋白，但不表达BMP-2、-5和-6的mRNA和蛋白。此外，基质细胞表达BMP I型受体、激活素样激酶-3（ALK-3）、ALK-6以及下游转导分子SMAD-1、-4和-5。因此，人骨髓基质细胞合成BMP，其可能通过特定的BMP受体以旁分泌方式对造血干细胞发挥作用。
