Human mesenchymal stem cells at the single-cell level: simultaneous seven-colour immunofluorescence.
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.
Schieker
Matthias
M
Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig-Maximilians-University (LMU) Munich, Germany. Matthias.Schieker@med.uni-muenchen.de
Pautke
Christoph
C
Haasters
Florian
F
Schieker
Jana
J
Docheva
Denitsa
D
Böcker
Wolfgang
W
Guelkan
Huelya
H
Neth
Peter
P
Jochum
Marianne
M
Mutschler
Wolf
W
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
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