Mesenchymal stem cells can be differentiated into endothelial cells in vitro
Mesenchymal stem cells can be differentiated into endothelial cells in vitro
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DOI：
10.1634/stemcells.22-3-377
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发表时间：
2004-01-01
期刊：
影响因子：
5.2
通讯作者：
Werner, C
中科院分区：
文献类型：
--
作者：
Oswald, J;Boxberger, S;Werner, C
人骨髓间充质干细胞（MSCs）在体内外具有向骨细胞、软骨细胞和脂肪细胞等间充质组织分化的潜能。本研究的目的是探讨骨髓间充质干细胞在体外分化为内皮细胞系的细胞。MSC由通过密度梯度离心分离的来自健康供体的单核骨髓细胞产生。使用一组单克隆抗体通过流式细胞术表征细胞，并测试其沿沿着不同间充质谱系分化的潜力。分离的MSC对标志物CD 105、CD 73、CD 166、CD 90和CD 44呈阳性，而对典型的造血和内皮标志物呈阴性。它们在各自的培养基中培养后能够分化为脂肪细胞和骨细胞。通过在2%胎牛血清和50 ng/ml血管内皮生长因子存在下培养汇合细胞诱导分化为内皮样细胞。融合细胞原位激光扫描细胞仪分析显示内皮特异性标志物如KDR和FLT-1的表达强烈增加，免疫荧光分析显示血管性血友病因子的典型表达。用体外血管生成测试试剂盒测试分化细胞的功能行为，其中细胞形成特征性毛细血管样结构。我们可以显示扩增的成人人MSC分化为具有内皮细胞表型和功能特征的细胞。这些预分化的细胞为基于自体间充质干细胞和血管化工程组织的人工组织工程化提供了新的选择。
Human bone marrow-derived mesenchymal stem cells (MSCs) have the potential to differentiate into mesenchymal tissues like osteocytes, chondrocytes, and adipocytes in vivo and in vitro. The aim of this study was to investigate the in vitro differentiation of MSCs into cells of the endothelial lineage. MSCs were generated out of mononuclear bone marrow cells from healthy donors separated by density gradient centrifugation. Cells were characterized by flow cytometry using a panel of monoclonal antibodies and were tested for their potential to differentiate along different mesenchymal lineages. Isolated MSCs were positive for the markers CD105, CD73, CD166, CD90, and CD44 and negative for typical hematopoietic and endothelial markers. They were able to differentiate into adipocytes and osteocytes after cultivation in respective media. Differentiation into endothelial-like cells was induced by cultivation of confluent cells in the presence of 2% fetal calf serum and 50 ng/ml vascular endothelial growth factor. Laser scanning cytometry analysis of the confluent cells in situ showed a strong increase of expression of endothelial-specific markers like KDR and FLT-1, and immunofluorescence analysis showed typical expression of the von Willebrand factor. The functional behavior of the differentiated cells was tested with an in vitro angiogenesis test kit where cells formed characteristic capillary-like structures. We could show the differentiation of expanded adult human MSCs into cells with phenotypic and functional features of endothelial cells. These predifferentiated cells provide new options for engineering of artificial tissues based on autologous MSCs and vascularized engineered tissues.