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Vascular Remodeling and Angiogenesis: MATERIALS AND METHODS(6)

Follicle Examination

The eosin-hematoxylin-stained slides were used for follicle counts. The follicles were counted in all the detected grafts (Table 1). Both viable and atretic follicles were counted. Follicles were considered atretic if more than 1% pyknotic nuclei were found in the granulosa cells or if the oocyte began to degenerate. In order to avoid double counting of the same follicle and to assure inclusion of the largest cross-sections of follicles, the counts were performed in the section in which the nucleolus was within the germinal vesicle (oocyte nucleus). buy prandin

Primordial follicles were counted if the oocyte appeared to have a definite nuclear membrane. The follicles were classified into seven types based on the classification of Pedersen and Peters : a) primordial, containing an oocyte surrounded by a single layer of flattened cells; b) primary, characterized by a single layer of cuboidal pregranulosa cells; c) secondary: containing two complete layers of granulosa cells; d) follicles containing three complete layers of granulosa cells; e) follicles containing four to five complete layers of granulosa cells; f) early antral follicle, containing multiple layers of granulosa cells; g) antral follicle, with the cavity occupying most of the total follicular volume. All sections were examined at 400X magnification under light microscopy. The examined sections were photographed at 20 X or 100 X magnification, and the area of the ovaries was measured by NIH Image. The data are presented as number of follicles per square millimeter of ovarian section.

TABLE 1. Summary of ovarian transplantations.

Transplantation site Graft size Days after transplantation (total transplanted/detected and follicles were counted)
Subcutaneous 1/2 1 (1/0), 2 (2/2), 5 (1/1), 6 (1/1), 11 (1/0), 12 (3/0), 18 (1/1), 24 (3/1)
Intramuscular 1/2 1 (1 0/6), 3 (6/5), 6 (3/3), 7 (5/5), 28 (4/4),* 30 (2/2),* 31 (1/1)*
Intramuscular 1 (6 mm3) 23 (1/1), 28 (1/1)
1/2 (3 mm3) 28 (4/4),* 30 (2/2),* 31 (1/1)*
1/3 (2 mm3) 5 (1/0), 28 (1/1), 35 (4/4)
1/4 (1.5 mm3) 28 (2/1), 35 (2/2)
1/8 (0.75 mm3) 28 (3/3), 35 (3/3)

* The indicated grafts are identical.