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Denney, N.

Publications and source records attributed to Denney, N..

2 recordsLinked to original sources

Optimizing the efficiency of assisted reproduction procedures in the common marmoset (Callithrix jacchus) improved IVF outcomes

The marmoset has emerged as a model species for genetic studies of complex neurodevelopmental conditions because of their brain development, behavior, communication, and compact size. IVF has been used to generate wild-type and transgenic marmoset embryos, but inefficiency of IVF in the marmoset compared to other primates poses challenges for genetic manipulation. To address this limitation, we analyzed the association between variables of age, number of cycles, and progesterone levels for IVF and embryology outcomes. Longitudinal data collected over 2 years indicated that the progesterone levels 24 hours after Cloprostenol sodium (Estrumate) administration, the number of days of ovarian stimulation during an IVF cycle, and the number of oocyte retrievals that an animal had undergone all influenced IVF cycle outcomes. Higher serum progesterone levels at cycle start were negatively correlated with the likelihood of fertilized eggs progressing to the cleavage stage of embryo development. Additionally, a shorter duration of ovarian stimulation (ranging 7 to 14 days of medication) coupled with a GnRH antagonist correlated with an increased percentage of fertilized eggs. We observed a trend that a shorter hormonal stimulation cycle length was associated with a higher percentage of embryos reaching the cleavage stage. The optimization of stimulation cycle protocol and embryology has led to a high percentage of implantation after embryo transfer regardless of whether the embryo was fresh or cryopreserved. These findings suggest that shorter IVF cycles and a progesterone level <10 ng/mL at cycle start were correlated with an increased likelihood of a cleavage stage embryo development.

developmental biology↗

Analysis of sperm processing and cryopreservation procedures in the common marmoset (Callithrix jacchus) to reduce fragmentation and maximize total and progressive motility

The marmoset has emerged as an ideal species for studying complex neurodevelopmental diseases, and IVF is required for gene-editing success. Reliable sperm processing is paramount to several IVF techniques, including traditional insemination. If not used immediately, thermal stress from cryopreservation diminishes sperm quality, requiring improved processing methods. Swim-up and gradient methods may limit adequate sperm quality, and the lack of a standardized freezing protocol often results in impaired post-thaw motility. Thus, reliable standards for sperm processing and cryopreservation in the marmoset are needed to support vital IVF processes. Collected semen underwent one of four pre-processing incubation steps before processing by swim-up, sucrose gradient, or a microfluidics assay. Sperm parameters were assessed using a Hamilton Thorne Computer Assisted Sperm Analysis (CASA) system. To evaluate cryopreservation, microfluidics-processed sperm were subjected to five protocols using different cryoprotectants and cooling methods. After thawing, sperm samples were analyzed with the CASA system and a DNA fragmentation assay. Incubation for 30 minutes at 38C in TYH media decreased sperm aggregation, and subsequent microfluidics resulted in a higher percentage of motile and progressively motile sperm than swim-up or gradient methods. Our optimized cryopreservation protocol consistently achieved at least 20% progressive motility, a key benchmark for usable marmoset sperm, while previously published or commercial methods were less consistent. The cryopreservation protocol outperformed other published methods, having the least significant change in post-thaw motility without causing DNA damage. These findings establish standardized methods for reliably procuring and storing sperm samples for IVF or husbandry in the common marmoset.

developmental biology↗