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Isolated follicles were more vulnerable to cryodamage after slow freezing as compared to vitrification.

The cytokine tumour necrosis factor-alpha (TNF alpha) has been postulated to play an essential role in the cytotoxic activity of cell-mediated immunity against allogenic or tumour cells invading the host. Several tumour cell lines, however, are resistant to TNF mediated cytotoxicity and respond paradoxically by cellular proliferation and by autocrine secretion of TNF alpha. In view of the metastatic character of the mammalian embryo, the aim of this study was to assess the potential of murine embryos to secrete TNF alpha in vitro, to express TNF receptors and to resist TNF alpha mediated cytotoxicity during their in-vitro development to the blastocyst stage. The potential of human embryos to secrete TNF alpha in vitro until the blastocyst stage was also investigated. From a total of 11 human embryos, which were allowed to proceed to blastocyst formation, seven secreted TNF alpha in the range of 2-117 pg/ml/24 h. A total of 123 C57BL/6J mouse embryos were studied of which 55% secreted TNF alpha in the range of 1.25-3.95 mg/ml/24 h. The presence of high levels of exogenous TNF alpha (10-300 IU) was not detrimental to the in-vitro development of murine embryos. Using immunohistochemical techniques, we were not able to detect the presence of type I or II TNF receptors on the surface of murine embryos. Our findings suggest that human and C57BL/6J murine embryos have the potential to secrete TNF alpha in vitro during the developmental stages leading to blastocyst formation.(ABSTRACT TRUNCATED AT 250 WORDS)

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