eb01198eadf38f4c81cb8eed51fb1be0 animals-15-00188_Podda_et_al_2025.pdf c56235635ae20f6d00a206265143ca4874b5cde6 animals-15-00188_Podda_et_al_2025.pdf 2d976d6b1e2d0b81a025c6b3feb5ef864238f0ba2cea03caf462acd04409e565 animals-15-00188_Podda_et_al_2025.pdf Title: Effect of Liquid Marble 3D Culture System on In Vitro Maturation and Embryo Development of Prepubertal Goat Oocytes Subject: Suboptimal culture conditions during in vitro maturation (IVM) affect oocyte developmental competence and the viability of the resulting embryo. Three-dimensional (3D) culture systems provide a more biologically appropriate environment compared to traditional two-dimensional (2D) cultures. The aim of this study was to evaluate the effect of liquid marble (LM) microbioreactors as a 3D culture system on IVM and the subsequent embryo development of prepubertal goat oocytes. The cumulus–oocyte complexes (COCs) recovered from prepubertal goat ovaries underwent IVM in drops under oil (the 2D system and the control group) and in the 3D LM system (the LM group). After IVM, oocytes were parthenogenetically activated and cultured until the blastocyst stage. The control and LM groups showed similar rates of nuclear maturation (52.17% and 44.12%) and blastocyst formation (10.64% and 10.10%). Reactive oxygen species and glutathione levels and the density of transzonal projections (TZPs) in oocytes did not differ between groups. The LM system increased mitochondrial activity and modified the organization of these organelles in the oocyte cytoplasm compared to the control group. The LM microbioreactor demonstrated the ability to improve the mitochondrial status of the oocytes and was not harmful for oocyte IVM and subsequent embryo development. Therefore, LM could be used as a 3D cost-effective culture system for the IVM of prepubertal goat oocytes. Keywords: 3D culture system; liquid marble; prepubertal goat; oocytes; embryo development Author: Andrea Podda, Linda Dujíčková, Federica Ariu, Giovanni Giuseppe Leoni, Dolors Izquierdo, Maria-Teresa Paramio and Luisa Bogliolo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Sun Jan 12 06:14:11 2025 CET ModDate: Mon Jan 13 08:02:55 2025 CET Custom Metadata: no Metadata Stream: yes Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4931414 bytes Optimized: yes PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- MIKRSI+VnURWPalladioL Type 1 Custom yes yes yes 545 0 CZIGNS+URWPalladioL-Roma Type 1 Custom yes yes yes 549 0 NBVNEI+URWPalladioL-Bold Type 1 Custom yes yes yes 552 0 DHTELU+URWPalladioL-Ital Type 1 Custom yes yes yes 555 0 UNSLQC+EURM10 Type 1 Builtin yes yes 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TrueType WinAnsi yes yes no 339 0 LOJHBP+PalatinoLinotype TrueType MacRoman yes yes no 341 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-01-24 02:41:54 CET RepresentationInformation: animals-15-00188_Podda_et_al_2025.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-01-22 07:42:22 CET Size: 4931414 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Linearized PDF PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Oocyte Collection and In Vitro Maturation Destination: subsection.2.1 Item: Title: Evaluation of Oocyte Nuclear Stage Destination: subsection.2.2 Item: Title: Assessment of Transzonal Projections Destination: subsection.2.3 Item: Title: Measurement of Reactive Oxygen Species (ROS) and Glutathione (GSH) Levels Destination: subsection.2.4 Item: Title: Evaluation of Mitochondrial Activity and Distribution Destination: subsection.2.5 Item: Title: Parthenogenetic Activation of Oocytes and Embryo Culture Destination: subsection.2.6 Item: Title: Assessment of Blastocyst Cell Number Destination: subsection.2.7 Item: Title: Statistical Analysis Destination: subsection.2.8 Item: Title: Results Destination: section.3 Children: Item: Title: Effect of LM System on Nuclear Maturation Destination: subsection.3.1 Item: Title: Effect of LM System on Transzonal Projections Density Destination: subsection.3.2 Item: Title: Effect of LM System on ROS and GSH Levels Destination: subsection.3.3 Item: Title: Effect of LM System on Mitochondrial Status Destination: subsection.3.4 Item: Title: Effect of LM System on Embryo Development and Blastocyst Cell Number After Parthenogenetic Activation Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: MIKRSI+VnURWPalladioL FontSubset: true FirstChar: 225 LastChar: 225 FontDescriptor: FontName: MIKRSI+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: ZFGZDQ+CMSY10 FontSubset: true FirstChar: 2 LastChar: 110 FontDescriptor: FontName: ZFGZDQ+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true ToUnicode: true Font: BaseFont: URYLRZ+URWPalladioL-BoldItal FontSubset: true FirstChar: 110 LastChar: 110 FontDescriptor: FontName: URYLRZ+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, -300, 1073, 935 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: CZIGNS+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 243 FontDescriptor: FontName: CZIGNS+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 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oocyte developmental competence and the viability of the resulting embryo. Three-dimensional (3D) culture systems provide a more biologically appropriate environment compared to traditional two-dimensional (2D) cultures. The aim of this study was to evaluate the effect of liquid marble (LM) microbioreactors as a 3D culture system on IVM and the subsequent embryo development of prepubertal goat oocytes. The cumulus–oocyte complexes (COCs) recovered from prepubertal goat ovaries underwent IVM in drops under oil (the 2D system and the control group) and in the 3D LM system (the LM group). After IVM, oocytes were parthenogenetically activated and cultured until the blastocyst stage. The control and LM groups showed similar rates of nuclear maturation (52.17% and 44.12%) and blastocyst formation (10.64% and 10.10%). Reactive oxygen species and glutathione levels and the density of transzonal projections (TZPs) in oocytes did not differ between groups. The LM system increased mitochondrial activity and modified the organization of these organelles in the oocyte cytoplasm compared to the control group. The LM microbioreactor demonstrated the ability to improve the mitochondrial status of the oocytes and was not harmful for oocyte IVM and subsequent embryo development. Therefore, LM could be used as a 3D cost-effective culture system for the IVM of prepubertal goat oocytes. 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