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Abstract Human fetal ovaries were obtained from second trimester foetuses (17 to 24 weeks gestation) after induced abortion. Ovaries were dissected from the fetal abdominal cavity and placed in collection medium for preservation. The tissues were cut into uniform sized wedges (3x 3x3 mm). Trypan blue solution was applied to ovarian samples to test its integrity. Any sample showing poor preservation was excluded from further processing. Ovarian fragments were frozen by using a cryopreservation protocol. Pieces of human fetal ovaries were thawed, fixed, embedded and sectioned at 5 J.lm thickness. The sections were deparaffinised and stained with Haematoxylin and Eosin staining. Human fetal ovaries were studied for the presence of germ cell markers using immunohistochemistry for STAT3, OCT4, VASA, active caspase-3 and Fas. Negative controls were performed by omitting the primary antibody which was replaced by normal serum resulting in complete absence of signals. The tissue fragments were cultured. Stem cell factor (SCF) was added to the medium. After 3 days and 7 days, individual cultures were prepared for analysis of cells using haematoxylin |