PubMed · 11649607
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Michael Gold. 1985. The baby makers.. https://pubmed.ncbi.nlm.nih.gov/11649607/
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Marine diplonemids (Diplonemidae and Hemistasiidae), once considered difficult to culture, can now be introduced into stable axenic cultures and grown to high densities. This chapter provides detailed protocols on establishing clonal cultures from environmental cells, their axenization and routine maintenance, as well as instructions on mid- and large-scale cultivation, estimation of cell density, and cryopreservation of selected species.
Oocyte cryopreservation would amount to a major breakthrough in reproductive medicine. Diverse strategies have been tested to minimise cooling-induced cell injury. Nevertheless, oocytes from various species have shown a particular sensitivity to freezing, due to their unique biological characteristics. Storage of human mature oocytes with slow freezing has resulted in low survival rates, although recent studies based on modified methods have reported higher success. Survival after thawing is not necessarily a guarantee of unaltered viability. Developmental failure at pre- or postimplantation stages may originate from critical perturbations of various cell components, such as the chromosome segregation apparatus, the intracellular calcium signalling system, and the cytoskeleton. Germinal vesicle (GV)-stage oocytes have been suggested to be more amenable to freezing. But their use would require efficient in vitro maturation systems, which are not presently available. Oocyte cryopreservation remains a realistic objective, provided that more systematic approaches are applied, such as thorough analysis of the plasmalemma permeability to water and the diverse cryoprotectants.
Couples accepting embryo cryopreservation signed an informed consent in which directives were given in case of death, divorce or absence. In this study, we tried to analyse our experience in terms of benefits achieved and to understand the feelings of couples about their embryos. (1) The majority of couples decided, or simply let, their embryos be discarded. In fact, a considerable proportion of them (25.1%) did not want to assume the responsibility of signing the disposal of their embryos. (2) Embryo donation is the most uncommon and difficult choice to make (6.0%) and this ought to be an important point to think over while dealing with law design. (3) An unexpected proportion of couples, who did not conceive, still have their embryos stored and this forces us to reflect on the difficulty of getting over the treatment failure.