Fig. 9
Transplantation of final hDPSC products from WCB improves CCl4-induced pro-fibrotic markers in mice. Two cryopreserved hDPSC products in WCB (donor #2 and #10) were randomly selected from the WCB batch and were thawed to use as final hDPSC products for the quality check. a Cell viability of the final hDPSC products. The final hDPSC products thawed from 10 vials. They were stained with trypan blue, and living cells were counted. Black columns, living cell numbers of the final hDPSC products at freezing; White columns, living cell number of the final hDPSC products after thawing. The graph bars show the mean ± SEM. n = 10. b, c Cell surface marker analysis of the final hDPSC products was tested by flow cytometric assay. Markers for mesenchymal stem cells (b) and immunogenic antigen (c) were assessed in the final hDPSC products. The results were shown as the averaged positive rates of each marker in each donor. n = 3. d–f Each final hDPSC products were intrasplenically transplanted into 4-week-CCl4-treated immunocompetent mice without immunosuppressants (n = 5). The age-matched control non-CCl4-treated (n = 5) and non-transplanted CCl4-treated (n = 5) immunocompetent mice were used as experimental controls. Serum levels of AST, ALT, total bilirubin (d), and human ALB (e) were examined by biochemical assays and ELISA. Distribution of the donor cells and deposition of fibrous tissues in the CCl4-damaged fibrotic liver tissues was investigated by immunohistochemical assay and collagen staining. The results are shown as representative images stained with antibodies against HLA-ABC, HepPar1, ACTA2, and Picrosirius Red. Nuclei are stained with hematoxylin. Bars = 50 μm (f). The liver fibrosis stages were assessed by Ishak scoring (g). d, e, hn = 5 for all groups. *P < 0.05, ***P < 0.005. The graph bars show the mean ± SEM (black columns) or the mean (white columns). ns, not significant