Figure 5.

Flow cytometric analysis of a live donor hepatocyte population from a 5-month-old hPAP transgenic donor. A: Acquisition dot plot showing three relatively distinct populations of viable hepatocytes. The gated subpopulations of hepatocytes were designated as R2, R3, and R4, corresponding to diploid, tetraploid, and octaploid, respectively (see text). Note that, as expected for a mouse of this age, the diploid cell population is a minority representing 5.6% of all gated cells. In contrast, tetraploid and octaploid cells accounted for 49% and 44% of the gated populations, respectively. The population of very small particles in the lower left-hand corner of the plot is thought to represent cellular debris. The population of very large cells to the right of R4 is highly enriched for cell doublets, as determined by microscopic examination of cells isolated from this region (data not shown). B: Acquisition histogram showing three distinct peaks. The M2, M3, and M4 populations correspond to the R2, R3, and R4 populations in the acquisition dot plot. Note that fluorescent areas of M3 and M4 are two and four times as large as the M2 population fluorescent area, respectively. C: Acquisition histogram of resorted live donor hepatocyte subpopulations. Top, middle, and bottom panels represent the resort of the original M2, M3, and M4 live hepatocyte subpopulations, respectively. Hoechst dye was re-added to the cell populations before the second sort, so that Hoechst fluorescent areas of the original and resorted cell populations are not precisely equivalent. During the resort, each population was collected separately and for a variable length of time, so the number of cells in each resort population is not proportional to the fraction of that population in the original sort.