Human mesenchymal stem cells at the single-cell level:: simultaneous seven-colour immunofluorescence

Human mesenchymal stem cells at the single-cell level:: simultaneous seven-colour immunofluorescence
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DOI:
10.1111/j.1469-7580.2007.00716.x
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发表时间:
2007-05-01
期刊:
影响因子:
2.4
通讯作者:
Mutschler, Wolf
Mutschler, Wolf
中科院分区:
医学3区
文献类型:
--
作者:
Schieker, Matthias;Pautke, Christoph;Mutschler, Wolf

文献摘要

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Extracellular, intracellular or surface proteins can be used as putative markers to characterize human mesenchymal stem cells (hMSC). However, these markers are also expressed by other cell types and primary cell pools reveal considerable heterogeneity. Therefore, the simultaneous detection of several markers on a single cell appears to be an attractive approach to identify hMSC. Here we demonstrate the specific distinction of human MSC from human osteoblasts via seven-colour fluorescence on the single cell level with simultaneous marker detection of CD44, CD105/endoglin, CD106/VCAM-1, collagen-IV, fibronectin, actin and DAPI nuclear staining. We performed spectral image acquisition using a Sagnac-type interferometer. Subsequent linear unmixing allowed for decomposition of each pixel in its spectral components. Our approach reveals a typical expression profile of the adherent singular cells, allowing the specific distinction between hMSC and osteoblasts on the single cell level.