Fig. 6.

Characterization of NG2⁺ cells isolated from human marrow MSCs (h-_BMMSCs) via the PPWP. (Aa-c) h-_BMMSC-sourced NG2⁺ cells (h-NG2/_BMMSCs) (a) isolated from h-_BMMSCs (b) and comparative analysis in size (c). The bold arrow indicates NG2⁺ cells, and the thin arrow indicates spindle-shaped _BMMSCs. (Ba, b) IF staining for Ki-67⁺ cells (red, arrows) in the two types of cells when their in normal cultures (a) and quantification (b); n = 3. (Ca-c) In normal _BMMSC cultures, larger flaky cells (a bold arrow, assumed to indicate NG2⁺ cells) appeared to produce smaller spindle-shaped cells (a thin arrow) (a, S1 and S2 represent individual cultures). IF staining for SSEA-3 (arrows, b) and comparative quantification of the two types of cells are shown (c); the box shows a cell clone. (Da-f) IF staining (red) of host CK19⁺ and Alb⁺ cells in the liver subgroups four weeks after cell transplantation (a, d), quantification (b, arrows; e, boxes)n = 6, and x-fold changes in host CK19⁺ cells stimulated by h-NG2/_BMMSCs compared with those stimulated by parental h-_BMMSCs (c, f). (Ea-f) IF staining (red) of host ɑ-SMA⁺ cells in DEN-induced mouse livers 4 weeks after transplantation of the two types of donor cells (b, c) compared to DEN (a) and quantification of protein (d, boxes, n = 6) and mRNA levels using RT‒qPCR (e, n = 6); blood functional hepatic parameters were also compared in these subgroups(f, n = 6). (Fa-f) IF staining (red) for host Lyve-1⁺ cells in subgroups of naive (a), DEN (b), two types of donor cells (c, d) four weeks after cell transplantation, and quantification of the number of Lyve-1⁺ cells per quarter area (boxes) of the staining (e, n = 6); mRNA levels in livers from the same subgroups were determined using RT‒qPCR (f, n = 6). Scale bar = 200 μm in all images; at least three independent experiments were performed, and the data are presented as the means ± SDs. *#p < 0.05 compared with naive or DEN or h-_BMMSCs; **p < 0.001 h-_BMMSCs; ns: indicate no significance