Isolation of Mouse Hepatic Stem Cells

Mouse hepatic stem cells were sorted from the liver of embryonic day 13.5 C57BL/6 fetal mice (n = 4), as we described previously [31 (link)]. In brief, liver cells were stained with the following antibodies: biotinylated anti-Ter-119 and anti-CD45, anti-c-Kit-APC, anti-CD49f-PE, and anti-CD29-FITC. Streptavidin-labeled allophycocyanin-Cy7 was used to detect biotinylated antibodies. All the above antibodies were purchased from BD Pharmingen, San Diego, CA, USA. For gating, the CD45⁻Ter119⁻ c-Kit⁻ cell population was firstly gated out, and then the CD49f^+/lowCD29⁺ subpopulation was set as the sorting gate. Analysis and sorting were performed using MoFlo with Summit version 4.0 software (DakoCytomation, Denmark).