Fig. 2
Self renewal assay for Hepa1–6 CSCs: Hepa1–6 CSC spheroids possess self-renewal capability in serum-free condition. Hepa1–6 cells were seeded in 96-well plate with appropriate dilution to achieve single cell/well, and observed for two weeks. a Representative bright field images of Hepa1–6 cells being self-renewed in serum free media at different time-points to form spheroids; b Fluorescence microscope images of growing spheroids at different time-points. DAPI (Blue) and FITC-EpCAM (CSC marker - Green) staining; c Bar graph showing data of Hepa1–6 cells being self-renewed to grow as spheroids; d Quantified cells for self-renewal (spheroid formation), differentiation (adherent growth), and single cell (quiescent cells). Images captured at 10× magnification (Bar = 200 μm)