Ploidy Reductions in Murine Fusion-Derived Hepatocytes (original) (raw)
Figure 3
Diploid hepatocytes express markers of cell fusion.
(A) Transplantation scheme for detection of fusion-derived hepatocytes. Donor bone marrow cells (from female transgenic mice hemizygous for R26R) were transplanted into lethally irradiated male Fah−/− mice containing the Alb-Cre transgene. Following fusion between donor hematopoietic cells and host hepatocytes, loxP sites were excised by Cre recombinase, leading to ß-gal expression via the ROSA26 promoter. (B and C) Hepatocytes from repopulated mice expressed ß-gal (blue, as seen by X-Gal staining) (B) and FAH (red) (C). Scale bars are 100 µm (n = 5). (D and E) Hepatocyte suspensions were loaded with Hoechst 33342 and FDG. Livers from repopulated mice were comprised of diploid and polyploid hepatocytes (D). FACS analysis shows the ploidy distribution from a representative mouse (n = 8). ß-gal was expressed by a subset of each hepatocyte ploidy population (E). FACS analysis shows FDG expression by diploid, polyploid and unfractionated hepatocytes from a representative liver (n = 5). (F) FAH was expressed by unfractionated hepatocytes as well as FACS-sorted diploid hepatocytes. Data represent percentage (average±SEM) of cells expressing FAH (n = 4).