Hematopoietic origins of fibroblasts: II. In vitro studies of fibroblasts, CFU-F, and fibrocytes
Hematopoietic origins of fibroblasts: II. In vitro studies of fibroblasts, CFU-F, and fibrocytes
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DOI：
10.1016/j.exphem.2005.10.008
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发表时间：
2006-02-01
影响因子：
2.6
通讯作者：
Ogawa, M
中科院分区：
文献类型：
--
作者：
Ebihara, Y;Masuya, M;Ogawa, M
Objective.通过移植来自转基因增强型绿色荧光蛋白（EGFP）小鼠单个造血干细胞（HSC）的克隆细胞群，我们已经证明了肌成纤维细胞（例如肾系膜细胞和脑小胶质细胞）的造血起源。因为成肌纤维细胞被认为是成纤维细胞的活化形式，所以我们测试了成纤维细胞来源于HSC的假设。将来源于EGFP Ly-5.2 C57 Bl/6小鼠的单细胞的细胞克隆移植到致死辐射的Ly-5.1小鼠中。利用小鼠的骨髓和外周血细胞显示高水平的多系造血重建，我们在体外诱导成纤维细胞的生长。从克隆移植的小鼠的EGFP（+）骨髓细胞培养显示贴壁细胞具有典型的成纤维细胞形态。流式细胞术分析显示，这些细胞中的大多数是CD 45（-），并表达胶原-I和胶原受体，盘状结构域受体2（DDR2）。培养细胞的逆转录聚合酶链反应分析显示前胶原1-α 1、DDR2、纤连蛋白和波形蛋白mRNA的表达。在来自克隆移植小鼠的骨髓细胞培养物中观察到由EGFP（+）细胞组成的成纤维细胞集落，表明成纤维细胞集落形成单位的HSC起源。克隆移植小鼠外周血有核细胞培养显示EGFP（+）细胞表达I型胶原和DDR2，提示纤维细胞也来源于HSC。我们得出结论，成纤维细胞和它们的前体细胞的人口来自HSC。(C)2006年国际实验血液学学会。爱思唯尔公司出版
Objective. Using transplantation of a clonal population of cells derived from a single hematopoietic stem cell (HSC) of transgenic enhanced green fluorescent protein (EGFP) mice, we have documented the hematopoietic origin of myofibroblasts, such as kidney mesangial cells and brain microglial cells. Because myotibroblasts are thought to be an activated form of fibroblasts, we tested the hypothesis that fibroblasts are derived from HSCs.Materials and Methods. Clones of cells derived from single cells of EGFP Ly-5.2 C57Bl/6 mice were transplanted into lethally irradiated Ly-5.1 mice. Using bone marrow and peripheral blood cells from mice showing high-level multilineage hematopoietic reconstitution, we induced growth of fibroblasts in vitro.Results. Culture of EGFP(+) bone marrow cells from clonally engrafted mice revealed adherent cells with morphology typical of fibroblasts. Flow cytometric analysis revealed that the majority of these cells are CD45(-) and express collagen-I and the collagen receptor, discoidin domain receptor 2 (DDR2). Reverse transcriptase polymerase chain reaction analysis of cultured cells demonstrated expression of procollagen 1-alpha 1, DDR2, fibronectin, and vimentin mRNA. Fibroblast colonies consisting of EGFP(+) cells were observed in cultures of bone marrow cells from clonally engrafted mice, indicating an HSC origin of fibroblast colony-forming units. Culture of peripheral blood nucleated cells from clonally engrafted mice revealed EGFP(+) cells expressing collagen-I and DDR2, indicating that fibrocytes are also derived from HSCs.Conclusion. We conclude that a population of fibroblasts and their precursors are derived from HSCs. (C) 2006 International Society for Experimental Hematology. Published by Elsevier Inc.