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Analysis of the purity of freshly isolated HSC obtained by density gradient centrifugation after plating and 1 day of culture. (A) Phase cytochrome c fragment (93-10 cool microscopic overview of a HSC primary culture with a purity of greater than 98%. (B) The typical cell morphology of rat HSC with perinuclear lipid droplets would allow the detection of other cell types. A small subset of floating HSC was still visible after 1 day of culture. (C) Immunofluorescence staining of the stellate cell marker glial fibrillary acidic protein (GFAP). (D) Detection of the stem/progenitor cell marker octamer binding factor 4 (Oct4) in the cell nuclei of HSC by immunofluorescence staining. The cell nuclei were labeled by DAPI staining to visualize all cells. The fluorescence pictures were taken by a confocal laser scanning microscope.
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Supplemental Fig. S3.
The expression of Wnt signaling elements was investigated in HSC cultured for 1 and 7 days by RT-PCR. Cells from the fetal rat brain (FRB; 20 days post coitum) served as a positive control. (A) Wnt ligands as well as their (B) receptors and co-receptors were analyzed. The mRNA of proteins of (C) the β-catenin destruction complex, (D) elements of Wnt signal transduction, and intracellular modulators of Wnt signaling were detected. (E) Transcription factors as well as their isoforms were also investigated on mRNA level. (F) The mRNA of extra cellular inhibitory proteins of Wnt signaling were found in HSC.