Intramuscular Ovary Transplantation: Optimization of Graft Size
Because the subcutaneous region is heterogeneous and relatively poor in its blood vessel support, we chose the muscle, which is more homogeneous and rich with vasculature, as a transplantation site. While the graft (in muscle as well as in the subcutaneous transplantations) should be large enough to contain the maximum pool of oocytes, it should also be small enough to minimize ischemia-reper-fusion injury. In order to optimize the graft size in the intramuscular transplantation, we studied various sizes of ovarian grafts, ranging from intact ovaries (6 mm3) to 1/8 ovary (0.75 mm3; Fig. 2).
The grafts were evaluated 2335 days after the transplantation. We detected 23 out of the 25 grafts (Table 1), and all of the detected grafts showed relatively good survival (low necrosis) and follicle maintenance. Normal appearance of blood vessels was observed using endothelial (Fig. 2, middle column) and smooth muscle cell markers (Fig. 2, right column). Follicles in different developmental stages were detected. No significant differences were observed between the average grades of the different graft sizes (data not shown). The follicles developed to the antral stage (stage g) only in the half ovarian grafts (3 mm3; Fig. 2, D-F). buy glycomet
FIG. 2. Ovary sections of various graft sizes from 15-day-old Wistar rats transplanted intramuscularly into CD-1 nude mice. Ovaries and ovary fragments were transplanted into the gluteus superficialis muscle at the hind limb. The grafts were retrieved 28-35 days after transplantation. A-C) Intact ovaries (6 mm3; n = 2). D-F) One-half ovary (3 mm3; n = 7), (G-I) 1/3 ovary (2 mm3; n = 5), (J-L) 1/4 ovary (1.5 mm3; n = 3), and (M-O) 1/8 ovary (0.75 mm3; n = 6). Left column: eosin hematoxylin staining; center column: endothelial cells staining (GSL-1) positive stain in brown; right column: smooth muscle cells and pericytes staining (aSMA) positive stain in red. Bar = 100 ^m.