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Human mesenchymal stem cells at the single‐cell level: simultaneous seven‐colour immunofluorescence
Author(s) -
Schieker Matthias,
Pautke Christoph,
Haasters Florian,
Schieker Jana,
Docheva Denitsa,
Böcker Wolfgang,
Guelkan Huelya,
Neth Peter,
Jochum Marianne,
Mutschler Wolf
Publication year - 2007
Publication title -
journal of anatomy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 118
eISSN - 1469-7580
pISSN - 0021-8782
DOI - 10.1111/j.1469-7580.2007.00716.x
Subject(s) - mesenchymal stem cell , cell , fibronectin , cell type , dapi , immunofluorescence , cd44 , extracellular , microbiology and biotechnology , biology , staining , chemistry , extracellular matrix , immunology , biochemistry , antibody , genetics
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.