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Albuquerque, NM, United States

Reed M.L.,Center for Reproductive Medicine of New Mexico
Methods in Molecular Biology | Year: 2012

Embryo density is defined as the embryo-to-volume ratio achieved during in vitro culture; in other words, it is the number of embryos in a defined volume of culture medium. The same density can be achieved by manipulating either the number of embryos in a given volume of medium, or manipulating the volume of the medium for a given number of embryos: for example, a microdrop with five embryos in a 50 μl volume under oil has the same embryo-to-volume ratio (1:10 μl) as a microdrop with one embryo in a 10 μl volume under oil (1:10 μl). Increased embryo density can improve mammalian embryo development in vitro; however, the mechanism(s) responsible for this effect may be different with respect to which method is used to increase embryo density. Standard, flat sterile plastic petri dishes are the most common, traditional platform for embryo culture. Microdrops under a mineral oil overlay can be prepared to control embryo density, but it is critical that dish preparation is consistent, where appropriate techniques are applied to prevent microdrop dehydration during preparation, and results of any data collection are reliable, and repeatable. There are newer dishes available from several manufacturers that are specifically designed for embryo culture; most are readily available for use with human embryos. The concept behind these newer dishes relies on fabrication of conical and smaller volume wells into the dish design, so that embryos rest at the lowest point in the wells, and where putative embryotrophic factors may concentrate. Embryo density is not usually considered by the embryologist as a technique in and of itself; rather, the decision to culture embryos in groups or individually is protocol-driven, and is based more on convenience or the need to collect data on individual embryos. Embryo density can be controlled, and as such, it can be utilized as a simple, yet effective tool to improve in vitro development of human embryos. © 2012 Springer Science+Business Media New York. Source


Said A.-H.,Center for Reproductive Medicine of New Mexico | Reed M.L.,Center for Reproductive Medicine of New Mexico
Journal of Assisted Reproduction and Genetics | Year: 2015

Purpose: The purpose of this study was to quantitate changes in seminal volume, sperm count, motility, qualitative forward progression, and total motile sperm cells per ejaculate, across three consecutive ejaculates collected from individuals within 24 h preceding an IVF cycle. Methods: Men presenting with oligoasthenozoospermia or asthenozoospemia attempted three ejaculates within 24 h preceding IVF. Ejaculate 1 was produced the afternoon prior to oocyte retrieval, and ejaculates 2 and 3 were produced the morning of oocyte retrieval with 2–3 h between collections. Ejaculates 1 and 2 were extended 1:1 v/v with room temperature rTYBS. Test tubes were placed into a beaker of room temperature water, then placed at 4 °C for gradual cooling. Ejaculate 3 was not extended, but pooled with ejaculates 1 and 2 and processed for intracytoplasmic sperm injection (ICSI). Results: Out of 109 oocyte retrievals, 28 men were asked to attempt multiple consecutive ejaculations. Among this population, 25/28 (89.3 %) were successful, and 3/28 men (10.7 %) could only produce two ejaculates. Mean volumes for ejaculates 1, 2, and 3 were significantly different from each other (p < 0.01); the volume decreased for each ejaculate. Mean sperm counts, motility, qualitative forward progression, and total motile cells per ejaculate for the ejaculates1, 2, and 3 demonstrated the following: ejaculates 2 and 3 were not significantly different, but counts, motility, and total motile sperm were improved over ejaculate 1 (p < 0.01). Conclusions: Pooling three consecutive ejaculates within 24 h increased the numbers of available motile sperm in this population by 8-fold compared to the first ejaculate alone, facilitating avoidance of sperm cryopreservation and additional centrifugation steps that could affect sperm viability and/or function. © 2015, Springer Science+Business Media New York. Source


Reed M.L.,Center for Reproductive Medicine of New Mexico | Said A.-H.,Center for Reproductive Medicine of New Mexico | Thompson D.J.,Center for Reproductive Medicine of New Mexico | Caperton C.L.,Center for Reproductive Medicine of New Mexico
Journal of Assisted Reproduction and Genetics | Year: 2015

Purpose: To evaluate the transition from a proven slow-cooling cryopreservation method to a commercial large-volume vitrification system for human blastocysts.Methods: Retrospective analysis of de-identified laboratory and clinical data from January 2012 to present date for all frozen embryo replacement (FET) cycles was undertaken. Cryopreservation of trophectoderm-biopsied or non-biopsied blastocysts utilized during this time period was logged as either slow-cooling, small-volume vitrification, or large-volume vitrification. Blastocyst survival post-warm or post-thaw, clinical pregnancy following FET, and implantation rates were identified for each respective cryopreservation method.Results: Embryo survival was highest for large-volume vitrification compared to micro-volume vitrification and slow-cooling; 187/193 (96.9 %), 27/32 (84.4 %), and 244/272 (89.7 %), respectively. Survival of biopsied and non-biopsied blastocysts vitrified using the large-volume system was 105/109 (96.3 %) and 82/84 (97.6 %), respectively. Survival for micro-volume biopsied and non-biopsied blastocysts was 16/30 (83.3 %) and 2/2 (100.0 %) respectively. Slow-cooling post-thaw embryo survival was 272/244 (89.7 %). Clinical pregnancy and implantation rates outcomes for non-biopsied embryos were similar between large-volume and slow-cooling cryopreservation methods, 18/39 (46.2 %) clinical pregnancy and 24/82 (29.3 %) implantation/embryo, and 52/116 (44.8 %) clinical pregnancy and 67/244 (27.5 %) implantation/embryo, respectively. Comparing outcomes for biopsied embryos, clinical pregnancy and implantation rates were 39/67 (58.2 %) clinical pregnancy and 50/105 (47.6 %) implantation/embryo and 4/16 (25 %) clinical pregnancy and 6/25 (24.0 %) implantation/embryo, respectively.Conclusions: The LifeGlobal large-volume vitrification system proved to be very reliable, simple to learn and implement in the laboratory. Clinically large-volume vitrification was as, or more effective compared to slow-cooling cryopreservation in terms of recovery of viable embryos in this laboratory. © 2014, Springer Science+Business Media New York. Source


Reed M.L.,Center for Reproductive Medicine of New Mexico | Hamic A.,Center for Reproductive Medicine of New Mexico | Caperton C.L.,Center for Reproductive Medicine of New Mexico | Thompson D.J.,Center for Reproductive Medicine of New Mexico
Fertility and Sterility | Year: 2010

Objective: To report a live birth after transfer of anonymously donated, twice-cryopreserved embryos that had been stored in liquid nitrogen for approximately 13.5 years. Design: Case report. Setting: A private assisted reproduction center. Patient(s): A 44-year-old recipient of donated cryopreserved embryos. Intervention(s): Anonymous donation of cryopreserved blastocysts for procreation. Main Outcome Measure(s): Live birth after thawing and replacement of re-cryopreserved blastocysts. Result(s): Fourteen pronuclear-stage embryos and four cleavage-stage embryos were cryopreserved during a primary IVF cycle. In two separate cycles, one cycle for the primary patient and a subsequent cycle for the first embryo donor recipient, the 18 embryos were thawed and grown to the blastocyst stage for transfer. Supernumerary blastocysts (n = 5) not replaced at either of these two thaw cycles were re-cryopreserved and subsequently donated to another embryo donor recipient. Five blastocysts survived the thaw and three were transferred, resulting in a live birth. The embryos were cryopreserved for a cumulative storage time of approximately 4,909 days (13.4 years). Conclusion(s): The longevity (viability) of cryopreserved embryos maintained in liquid nitrogen remains to be determined; cryopreserved embryo donation for procreation should not be overlooked, regardless of the length of time that embryos remain in cryostorage. Copyright © 2010 American Society for Reproductive Medicine, Published by Elsevier Inc. Source

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