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New Time-Lapse Incubator for assisted reproduction research

The ICTS NANBIOSIS has expanded its capabilities with the installation and commissioning of a new equipment Time-Lapse Incubator in the CCMIJU’s Assisted Reproduction Lab.

The acquisition of the Time-Lapse Incubator is part of the project “Embryonic Genetics in Assisted Reproduction” (GENERA), co-funded by the European Regional Development Fund (ERDF) within the framework of Spain’s Plurirregional Operational Program for Singular Scientific and Technical Infrastructures (ICTS) 2014 -2020 and by Consejería de Economía, Ciencia y Agenda Cultural of  Junta de Extremadura.

The objective of GENERA includes the purchase of lab equipment to expand services in the field of embryonic genetics as well as creating, editing and making traceability of embryos with high genetic value.

The first lab equipment acquired is the EPPENDORF PiezoXpert Piezoelectric-assisted micromanipulator that supports the creation and possible embryo editing, allowing easy penetration into cells for subsequent microinjection or micromanipulation.

The second one is the Time-Lapse Incubator that enables observation of embryos for accurate assessments and minimizing embryo culture stress.

The purchase of this lab equipment will offer the possibility of developing next-generation embryos, being able to face specific studies of the highest reproductive level and offering studies demanded by companies in the sector.

The project has an eligible budget of €98,000, of which the ERDF co-financing rate (80%) amounts to 78,400 and the national contribution to €21,600. It is expected to be completed in June 2023.

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Seminal plasma influences the fertilizing potential of cryopreserved stallion sperm

Beatriz Macías García, researcher at  NANBIOSIS U23. Assisted Reproduction,  is co-author of the article “Seminal plasma influences the fertilizing potential of cryopreserved stallion sperm”, publish by the Journal Theriogenology.

 

Seminal plasma (SP) contains proteins that may influence cryosurvival and prevent capacitation-like changes due to freezing and thawing. The objective of this study was to investigate the effect of adding pooled SP from “good” (GF) or “bad” (BF) freezer stallions on sperm cells’ fertilizing ability. “Good freezers” refers to stallions that usually produce ejaculates which can withstand cryopreservation, whilst “bad freezer” stallions produce ejaculates which cannot tolerate the freezing process. A heterologous zona binding assay with in vitro matured bovine oocytes was used to assess the binding ability of equine sperm cells as a possible alternative to artificial insemination trials. The effect of adding SP i) prior to cryopreservation; ii) after thawing of sperm cells selected by single layer centrifugation (SLC); iii) to capacitation medium, was evaluated. Adding SP from GF stallions prior to cryopreservation reduced the mean number of sperm cells bound to the zona pellucida (ZP) compared to control (P = 0.0003), SP-free sperm cells and group received SP from BF stallions (P ≤ 0.0001 for both). After thawing SLC-selected sperm cells treated with 5% SP showed a decrease in binding ability compared with SP-free sperm cells (P ≤ 0.0001). The binding affinity of sperm cells was higher in the group treated with SP from GF than with SP from BF stallions (P ≤ 0.05). Prolonged exposure to SP impaired the ability of stallion sperm cells to undergo capacitation and bind to ZP, regardless of the source of SP (P ≤ 0.0001). The response of equine sperm cells to SP is influenced by the ability of the sperm cells to withstand cryopreservation and is affected by the timing of exposure and the origin of SP. Customization of the protocol for individual stallions is recommended to optimize the effect.

 

The research has been carried out with the participation of the NANBIOSIS  Assisted Reproduction Lab which ofer the services of

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Mimicking physiological O2 tension in the female reproductive tract improves assisted reproduction outcomes in pig

Francisco M. Sánchez-Margallo, Assistant Director of NANBIOSIS and Scientific Coordinator o NANBIOSIS U23. Assisted Reproduction,  is co-author of the article “Mimicking physiological O2 tension in the female reproductive tract improves assisted reproduction outcomes in pig”, publish by the Journal Molecular human reproduction

The research has been carried out with the participation of the NANBIOSIS  Assisted Reproduction Lab that has  a 120 m2 laboratory, small animal surgery, clinical analysis support, etc. and it is featured with two intracitoplasmatic micromanipulation equipment of the latest generation with IMSI, Laser and Oosight system, embryo biopsy systems, vision systems of the mitotic spindle, with flow cabinets with stereo-microscopes and heated plates, incubators with different gasses systems, equipment and cryopreservation freeze gamete and embryo, among others

STUDY QUESTION:

Is O2 tension in the pig oviduct and uterus affected by the estrous cycle stage and the animal’s age, and can the outcome of in vitro embryo development be improved by mimicking these physiological values?

SUMMARY ANSWER:

O2 tension within the pig reproductive organs is affected by the animal’s age, and values close to those measured in vivo have a positive impact on embryo development and quality when used during IVF and embryo culture (EC).

Article of reference: doi: 10.1093/molehr/gay008

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Relevant research results for assisted reproduction.

Javier García Casado, Scientific Director of NANBIOSIS Unit 14: Cell therapy toguether with other Scientists of NANBIOSIS and CCMIJU have issued the first report describing the beneficial effect of human EV-endMSCs on embryo development, which has been recently published by PloS One: “Murine embryos exposed to human endometrial MSCs-derived extracellular vesicles exhibit higher VEGF/PDGF AA release, increased blastomere count and hatching rates“.  The results could be relevant for assisted reproduction:

“Endometrial Mesenchymal Stromal Cells (endMSCs) are multipotent cells with immunomodulatory and pro-regenerative activity which is mainly mediated by a paracrine effect. The exosomes released by MSCs have become a promising therapeutic tool for the treatment of immune-mediated diseases. More specifically, extracellular vesicles derived from endMSCs (EV-endMSCs) have demonstrated a cardioprotective effect through the release of anti-apoptotic and pro-angiogenic factors. Here we hypothesize that EV-endMSCs may be used as a co-adjuvant to improve in vitro fertilization outcomes and embryo quality. Firstly, endMSCs and EV-endMSCs were isolated and phenotypically characterized for in vitro assays. Then, in vitro studies were performed on murine embryos co-cultured with EV-endMSCs at different concentrations. Our results firstly demonstrated a significant increase on the total blastomere count of expanded murine blastocysts. Moreover, EV-endMSCs triggered the release of pro-angiogenic molecules from embryos demonstrating an EV-endMSCs concentration-dependent increase of VEGF and PDGF-AA. The release of VEGF and PDGF-AA by the embryos may indicate that the beneficial effect of EV-endMSCs could be mediating not only an increase in the blastocyst’s total cell number, but also may promote endometrial angiogenesis, vascularization, differentiation and tissue remodeling”.

Soluble factors were analyzed by the ICTS Nanbiosis (Unit 14. Cell therapy at CCMIJU). Maintenance of animals was performed by the ICTS Nanbiosis (Unit 22. Animal housing at CCMIJU). In vivo embryo recovery and culture was performed by the ICTS Nanbiosis (Unit 23. Assisted Reproduction at CCMIJU)

Article of reference:

Blázquez R, Sánchez-Margallo FM, Álvarez V, Matilla E, Hernández N, Marinaro F, Gómez-Serrano M, Jorge I, Casado JG, Macías-García B. “Murine embryos exposed to human endometrial MSCs-derived extracellular vesicles exhibit higher VEGF/PDGF AA release, increased blastomere count and hatching rates”. PLoS One (2018-04; vol. 13(4))

DOI: 10.1371/journal.pone.0196080

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