Flow cytometry sorting pathway to purify the mature, early and committed progenitors of ICC.

<p>Analysis following gating: (<b>A</b>) Selection of living mononuclear cells on the histogram with Side Scatter (SSC)/Forward Scatter (FSC). R1 gate was used to select the cells with light scatter properties characteristic of live cells. (<b>B</b>) R2 gate was used to select the cells not expressing macrophage markers (F4/80, CD11b) and the general hematopoietic marker CD45 (FITC- cells): gating on histogram SSC/CD45. (<b>C</b>) Detection of ICC phenotypes: the c-Kit⁺ cells population was gated in R4 and further analyzed in step <b>E</b>; the <b>c-</b>Kit^low cells population was gated in R3 and further analyzed in next step <b>D</b>. (<b>D</b>) The cells in R5 were <b>c-</b>Kit^lowCD34⁺Igf1r⁺. (<b>E</b>) The cells in R6 were <b>c-</b>Kit⁺CD34⁺Igf1r⁺, and the cells in R7 were c-Kit⁺CD34⁻Igf1r⁻. (<b>F</b>) To confirm the presence of the c-Kit^low population (R3) reliably between c-Kit⁻ and c-Kit⁺ populations (R4). Pink discontinuous straight line was used as the dividing line between c-Kit⁻ and c-Kit^low populations.