BACKGROUND: The complexity of fertilization failure during assisted reproductive technologies (ART) is often under-appreciated, as this failure can occur at any number of essential mechanistic and cellular events. Importantly, successful fertilization is heavily dependent upon inherent qualities of the oocytes, and thus reliant upon fidelity of oocyte maturation. METHODS: Pubmed and medline were searched up to April 2008 for papers on oocyte fertilization and its mechanistic components. References to clinical/human studies were selected wherever possible. RESULTS: Successful oocyte maturation cannot simply be determined via visual assessment of polar body extrusion, but rather entails coordination of numerous cytoplasmic processes not readily observed. Proper regulation of intraoocyte signaling cascades is crucial for sufficient production and storage of carbohydrates and proteins, successful relocation of organelles and regulation of metabolic pathways required for an apparently mature metaphase II oocyte to complete subsequent fertilization events; such as cumulus penetration, sperm/oocyte binding, fusion, oocyte activation, sperm processing and pronuclear (PN) formation. Regulation of oocyte maturation begins during oocyte growth and is intimately connected with events influencing folliculogenesis. Therefore, the oocyte is subject to a multitude of potential effector impacting fertilization potential and developmental competence long before encountering the artificial environment of the IVF laboratory. CONCLUSIONS: Although meticulous care and continued research is essential for future improvement, failure to fertilize and properly form PN following clinical ART is likely to be dependent on historical events in oocyte maturation, not easily explained or prevented through simple modification of contemporary laboratory protocols.
Over the past decade there has been a resurgence of interest in the culture media used in clinical in vitro fertilization. Unfortunately, during this time more confusion than consensus appears to have developed regarding the composition of these media. In order to facilitate a clearer understanding of this field, it is important to understand the role of specific medium components and how their use is regulated by the embryo. The roles of the key nutrients glucose, pyruvate, lactate, and amino acids during the preimplantation period have therefore been presented. Analysis of how the embryo regulates the utilization of such nutrients has led to a clearer understanding of the embryo's requirements during the dynamic period of preimplantation development. From such information, sequential culture media have been developed along with novel noninvasive tests of embryonic viability. It is proposed that continued studies on the human embryo will lead to further improvements in embryo culture conditions and the optimization of viability assays, culminating in the ability to transfer single embryos for the majority of, if not all patients.
Maintenance of stable pH is important for optimizing gamete and embryo culture. One method to stabilize pH entails using zwitterionic buffers in IVF handling media used outside the laboratory incubator. Current handling media utilize single buffers, such as MOPS or HEPES. However, the use of a single buffer limits the ability to adjust the range of buffering capacity. Furthermore, changes in temperature alter buffering of these compounds. Therefore, traditional IVF handling media utilizing a single buffer may not provide ideal pH buffering. This study reports that combining multiple buffers, such as HEPES, MOPS and DIPSO, into a single medium in various ratios gives the ability to shift the effective buffering range to cover a specific pH. Additionally, by combining various buffers, it is possible to expand pH buffering over a range of temperatures, while simultaneously reducing the absolute concentration of individual buffers, thereby reducing or alleviating toxicity concerns. This report verifies that DIPSO, MOPS and HEPES, and their combinations, support embryo development. Therefore, utilization of bi- and tri-buffered media, containing a mixture of HEPES, MOPS or DIPSO, offers advantages compared with media containing HEPES or MOPS alone, and may be used for procedures such as oocyte retrieval, intracytoplasmic sperm injection, embryo transfer and cryopreservation.
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