Figure 8

Fate of exogenous GFP-positive follicles transplanted into recipient ovaries 1 week after B/C treatment.

(a) GFP expression vector. (b and c) GFP transgenic mice and GFP-positive cells, including primordial follicles, isolated from postnatal day 5 ovaries of GFP transgenic mice. (d) Transplantation of the GFP-positive female germ cells into ovaries at 1 week after B/C treatment. Inset in (d) depicts primordial follicles used for ovarian transplantation. Most of DAPI positive cells showed GFP positive. DAPI staining (e) and localization of exogenous- and endogenous-derived primordial follicles by using GFP expression and MVH staining soon after female germ cell transplantation (f). Both of MVH staining and GFP expression indicate these germ cells are derived from transplanted primordial follicles, whereas MVH staining or GFP expression alone means endogenous primordial follicle or exogenous derived somatic cells (exoS), respectively. Exogenous derived GFP positive primordial follicles were localized along with host follicles containing GFP-negative follicles (MVH positive) in ovaries of all female mice that received GFP-expressing primordial follicles (f). At 1 day post-transplantation, many of the exogenous GFP-positive female germ cells were found along the injection tracts (g). Based on observations from whole-mount preparations, however, some GFP-positive cells were scattered within the ovaries. At 1 week post-transplantation, the number of GFP-positive cells significantly decreased (h) and, by 5^th weeks, the GFP-positive cells disappeared (i). These time-lapse experiments showed that after transplantation, transplanted GFP-positive cells did not survive the effects of B/C toxicity. enF and exG indicate endogenous follicles and exogenous GFP-positive cells, respectively.