title: Human mesenchymal stem cells at the single-cell level: simultaneous seven-colour immunofluorescence.
abstract: 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.
authors: Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig-Maximilians-University (LMU) Munich, Germany. Matthias.Schieker@med.uni-muenchen.de
authors: Matthias
authors: M
authors: Schieker
authors: Christoph
authors: C
authors: Pautke
authors: Florian
authors: F
authors: Haasters
authors: Jana
authors: J
authors: Schieker
authors: Denitsa
authors: D
authors: Docheva
authors: Wolfgang
authors: W
authors: Böcker
authors: Huelya
authors: H
authors: Guelkan
authors: Peter
authors: P
authors: Neth
authors: Marianne
authors: M
authors: Jochum
authors: Wolf
authors: W
authors: Mutschler
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000032:analysis; D000071063:Endoglin; D005353:Fibronectins / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis; D007090:Image Interpretation, Computer-Assisted; D007091:Image Processing, Computer-Assisted; D059630:Mesenchymal Stem Cells / Q000737:chemistry / Q000166:cytology*; D010006:Osteoblasts / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000032:analysis; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D024141:Collagen Type IV; C064206:ENG protein, human; D000071063:Endoglin; D005353:Fibronectins; D005456:Fluorescent Dyes; D018960:Hyaluronan Receptors; D011956:Receptors, Cell Surface; D019010:Vascular Cell Adhesion Molecule-1