Transplanted HPLCs survived in the liver and produced human Factor IX. (a) HPLCs on Day 13 were labeled with CM-DiI. The red fluorescence showed the CM-DiI labeled cells. Scale bar: 500 µm. (b) Four weeks after intrasplenic transplantation of HPLCs (Red) into SCID mouse with carbon tetrachloride-induced liver damage. Immunofluorescence staining on liver sections was performed to detect human Factor IX (Green). Scale bar: 10 µm. (c) Four weeks after transplantation, the plasma of mice was collected and human Factor IX antigen was detected by ELISA. Blank, mice injected with PBS; U-HPLC, HPLCs differentiated from un-targeted mESCs; E2-HPLC, HPLCs differentiated from Neo cassette excised clone E2. ns, not significant; ***, p < 0.001.