Figure 2.
HPCs Have a Potential to Persistently Produce Myofibroblasts in Clonal Subcultures
(A) Experimental procedure to perform subcloning experiments for three different HPC clones (HPC-C1, HPC-C2, and HPC-C3). Cells in cultures of each HPC clone underwent clone sorting by flow cytometry and were clonally cultured in 96-well plates. After LC formation and clonal expansion, three representative subclones of the individual primary HPC clones were chosen for examination.
(B and C) The numbers of LCs in the wells of 96-well plates (B) and the numbers of LCs containing α-SMA+ cells in LCs (C) were counted. The percentages are shown.
(D) Immunofluorescence staining of α-SMA and co-immunofluorescence staining of ALB with α-SMA and CK19 were conducted for cells in LCs formed from cells in cultures of a primary HPC clone. DNA was stained with DAPI. Scale bars, 0.5 mm (left panel) and 50 μm (right panels).
(E) The percentages of cells immunoreactive for α-SMA among subclones of the three primary HPC clones were calculated after counting ∼1 × 105 cells in individual culture dishes. The data represent means ± SD of three technical replicates (n = 3).