Conditioned Medium from Human Amnion-Derived Mesenchymal Stem Cells Regulates Activation of Primary Hepatic Stellate Cells

Figure 2

Gating strategy for HSC purification using FACS and HSC characterization. (a) Cells were gated on the basis of their FSC and SSC, viable cells were selected using 7-AAD, and doublets were excluded by FSC-A/FSC-H. Finally, HSCs were selected on the basis of the cyan light emission from retinol. (b) Purity of HSCs after FACS. (c) Primary HSCs are irregularly round-shaped, and retinol-rich lipid droplets emit cyan autofluorescence by a 352 nm laser. The HSCs cultured for 2 days became extended and presented an asteroid phenotype, whereas large numbers of lipid droplets were observed still. No autofluorescence could be detected after culturing HSCs for 4 days, and the cells extended further. FSC: forward scatter; SSC: sideward scatter; 7-AAD: 7-aminoactinomycin D; NUV: near ultraviolet. Scale bar = 50 μm.

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