“…Many groups worldwide have achieved blastocyst rates similar to the ones reported here (∼40 %), indicating that co-culture with stem cells was as efficient as current IVEP protocols in bovines, most of them using supplemented IVC medium [10,[50][51][52][53][54]. Therefore, further identification of molecules produced by stem cells for embryo development could help to improve IVEP not only in bovine but also in other species such as humans.…”
Purpose Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development. Materials and methods In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 10 3 and 10 4 cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 10 4 b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran). Results In experiment 1, co-culture with 10 4 b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 10 3 b-ATMSCs/mL (p = 0.051), group 10 4 bATMSCs/mL yielded higher results of blastocyst production.In experiment 2, when compared to group Gran, co-culture with 10 4 b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05). Conclusions Co-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in bovine and other species.
“…Many groups worldwide have achieved blastocyst rates similar to the ones reported here (∼40 %), indicating that co-culture with stem cells was as efficient as current IVEP protocols in bovines, most of them using supplemented IVC medium [10,[50][51][52][53][54]. Therefore, further identification of molecules produced by stem cells for embryo development could help to improve IVEP not only in bovine but also in other species such as humans.…”
Purpose Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development. Materials and methods In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 10 3 and 10 4 cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 10 4 b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran). Results In experiment 1, co-culture with 10 4 b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 10 3 b-ATMSCs/mL (p = 0.051), group 10 4 bATMSCs/mL yielded higher results of blastocyst production.In experiment 2, when compared to group Gran, co-culture with 10 4 b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05). Conclusions Co-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in bovine and other species.
“…However, a growing number of studies have applied nanotechnology in reproductive biology to target and sort specific populations of gametes and to optimize gene transfer and editing methodologies (Barkalina, Charalambous, Jones, & Coward, ; Remião et al, ). In addition, nanoparticulated systems have shown great potential as carriers to deliver molecules into gametes and embryos to improve their developmental competence in vitro (Barkalina, Charalambous, et al, ; Barkalina et al, ; Komninou et al, ; Lucas et al, ; Remião et al, ).…”
Section: Nanotechnologymentioning
confidence: 99%
“…In addition, nanoencapsulation allowed for the use of lower concentrations of TTN, minimizing the risk of cytotoxic effects related with its use (Lucas et al, ). Similarly, melatonin‐loaded lipid‐core nanocapsules (Mel‐LNC) added during bovine IVM and IVC reduced ROS levels and apoptosis in embryos, increased the cleavage and blastocyst rates, upregulated antioxidant genes, and downregulated proapoptotic genes when compared to nonencapsulated melatonin (Mel) (Komninou et al, ; Remião et al, ). LNCs synthesized by using the Rhodamine B fluorophore–PCL conjugated to label the nanocarrier (Mel‐LNC‐RB) achieved successful delivery into bovine oocytes and remained inside the cells until the blastocyst stage (Remião et al, ).…”
Section: Nanotechnologymentioning
confidence: 99%
“…Similarly, melatonin‐loaded lipid‐core nanocapsules (Mel‐LNC) added during bovine IVM and IVC reduced ROS levels and apoptosis in embryos, increased the cleavage and blastocyst rates, upregulated antioxidant genes, and downregulated proapoptotic genes when compared to nonencapsulated melatonin (Mel) (Komninou et al, ; Remião et al, ). LNCs synthesized by using the Rhodamine B fluorophore–PCL conjugated to label the nanocarrier (Mel‐LNC‐RB) achieved successful delivery into bovine oocytes and remained inside the cells until the blastocyst stage (Remião et al, ). Recently, a 3D culture system, based on magnetic levitation with precise assembly of nanoparticles promoted the in vitro development of bovine secondary follicles.…”
An appropriate environment to optimize porcine preimplantation embryo production in vitro is required as genetically modified pigs have become indispensable for biomedical research and agriculture. To provide suitable culture conditions, omics technologies have been applied to elucidate which metabolic substrates and pathways are involved during early developmental processes. Metabolomic profiling and transcriptional analysis comparing in vivo‐ and in vitro‐derived embryos have demonstrated the important role of amino acids during preimplantation development. Transcriptional profiling studies have been helpful in assessing epigenetic reprogramming agents to allow for the correction of gene expression during the cloning process. Along with this, nanotechnology, which is a highly promising field, has allowed for the use of engineered nanoplatforms in reproductive biology. A growing number of studies have explored the use of nanoengineered materials for sorting, labeling, and targeting purposes; which demonstrates their potential to become one of the solutions for precise delivery of molecules into gametes and embryos. Considering the contributions of omics and the recent progress in nanoscience, in this review, we focused on their emerging applications for current in vitro pig embryo production systems to optimize the generation of genetically modified animals.
“…Enhances in vitro embryo production, decreases ROS levels and the apoptotic nuclei, upregulates GPX1 and SOD2 and downregulates CASP3 and BAX [81] Bovine Zygotes Melatonin-loaded lipid-core nanocapsules at 10 -9 M during in vitro culture…”
Oxidative and nitrosative stress are a common problem when manipulating gametes in vitro. In vitro development in mammalian embryos is highly affected by culture conditions, especially by reactive oxygen species (ROS) and reactive nitrogen species (RNS), because its absence or over production causes embryo arrest and changes in gene expression. Melatonin in gamete co-incubation during IVF has deleterious or positive effects depending on the concentration used in culture medium, demonstrating the delicate balance that must exist between antioxidant and pro-oxidant activity. Further research is needed to better understand the possible impact of melatonin on the different IVP steps in domestic animals, especially in seasonal breeds where this neuro-hormone system highly regulates its reproduction physiology.
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