Role of Tumor Associated Fibroblasts in Human Liver Regeneration, Cirrhosis, and Cancer

Figure 3

Multipotency of MASCs obtained from neoplastic livers. A neuroectodermic differentiation: N-MASCs (a) in neurogenic medium acquired the expression of β3-tubulin (green fluorescence). (b–e) Osteogenic differentiation: N-MASCs in osteogenic medium display positivity for von Kossa (b) and Alcyan blue (c) cytochemistry reactions, were able to incorporate tetracycline in sites of calcification when excited with a UV laser (red fluorescence, (d)) and presented osteocalcin deposits (green fluorescence, (e)). (f, g) Myocyte differentiation: a low fraction of N-MASCs in differentiation medium express the myocyte-specific filament SMA (green fluorescence, (f)), while the large part of the cells expresses α-sarcomeric actin in the cytoplasm (red fluorescence, (g)). (h–k) Endodermic differentiation: cells cultured for one week in a medium added with HGF and FGF-4 became positive for GATA-4 (yellow fluorescence, (h)). Cells cultured for three weeks in the same medium became positive for cytokeratins 8–18-19 (green fluorescence, (i)) were able to actively uptake DiI-labeled acetylated LDL (red fluorescence overlaid on a phase contrast image, (j)) and presented diastase-resistant glycogen granules, as revealed by PAS reaction (purple stain, (k)). In the fluorescence pictures, nuclei are depicted by the blue fluorescence of DAPI staining.