0d7b4077b1a673b8ba4119880896df09 ijmolsci_a2022v23n13p7069.pdf 83e7546637ea8eb2c68050d78fff32bca37b7890 ijmolsci_a2022v23n13p7069.pdf 8431c44ee9eb3af31a3b889e6970340d645c853f3a0b53a83143b50dc97d9549 ijmolsci_a2022v23n13p7069.pdf Title: Exopolysaccharide ID1 Improves Post-Warming Outcomes after Vitrification of In Vitro-Produced Bovine Embryos Subject: This study aimed to assess the cryoprotectant role of exopolysaccharide (EPS) ID1, produced by Antarctic Pseudomonas sp., in the vitrification of in vitro-produced (IVP) bovine embryos. IVP day 7 (D7) and day 8 (D8) expanded blastocysts derived from cow or calf oocytes were vitrified without supplementation (EPS0) or supplemented with 10 g/mL (EPS10) or 100 g/mL (EPS100) EPS ID1. The effect of EPS ID1 was assessed in post-warming re-expansion and hatching rates, differential cell count, apoptosis rate, and gene expression. EPS100 re-expansion rates were significantly higher than those observed for the EPS0 and EPS10 treatments, regardless of culture length or oocyte source. EPS100 hatching rate was similar to the one of the fresh blastocysts except for those D7 blastocysts derived from calf oocytes. No differences were observed among EPS ID1 treatments when the inner cell mass, trophectoderm, and total cell number were assessed. Although apoptosis rates were higher (p 0.05) in vitrified groups compared to fresh embryos, EPS100 blastocysts had a lower number (p 0.05) of apoptotic nuclei than the EPS0 or EPS10 groups. No differences in the expression of BCL2, AQP3, CX43, and SOD1 genes between treatments were observed. Vitrification without EPS ID1 supplementation produced blastocysts with significantly higher BAX gene expression, whereas treatment with 100 g/mL EPS ID1 returned BAX levels to those observed in non-vitrified blastocysts. Our results suggest that 100 g/mL EPS ID1 added to the vitrification media is beneficial for embryo cryopreservation because it results in higher re-expansion and hatching ability and it positively modulates apoptosis. Keywords: cryopreservation; blastocyst; total cell number; inner cell mass; TUNEL; embryo development; gene expression regulation Author: Erika Alina Ordóñez-León, Iris Martínez-Rodero, Tania García-Martínez, Manel López-Béjar, Marc Yeste, Elena Mercade and Teresa Mogas Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Jun 29 05:01:27 2022 CEST ModDate: Wed Jun 29 05:50:40 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1441925 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 XAWNGI+VnURWPalladioL Type 1 Custom yes yes no 15 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 19 0 ZTEVSZ+URWPalladioL-Bold Type 1 Custom yes yes no 23 0 SEUERA+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 HWQYTH+URWPalladioL-Ital Type 1 Custom yes yes no 32 0 IFEDSM+CMSY10 Type 1 Builtin yes yes no 36 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 54 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 57 0 URYLRZ+URWPalladioL-BoldItal Type 1 Custom yes yes no 82 0 MDIKPC+CMBSY10 Type 1 Builtin yes yes no 92 0 BGCFBF+CalibriLight CID TrueType Identity-H yes yes yes 105 0 BGCAAK+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 111 0 BGCAAL+PalatinoLinotype CID TrueType Identity-H yes yes yes 114 0 BGCAAM+PalatinoLinotype TrueType WinAnsi yes yes no 120 0 BGCAAN+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 123 0 BGCAAO+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 129 0 BGCAAK+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 158 0 BGCAAL+PalatinoLinotype CID TrueType Identity-H yes yes yes 161 0 BGCAAM+PalatinoLinotype TrueType WinAnsi yes yes no 167 0 BGCAAO+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 170 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 177 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-07-21 02:36:08 CEST RepresentationInformation: ijmolsci_a2022v23n13p7069.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-06-29 10:48:30 CEST Size: 1441925 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 391 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Chemicals and Suppliers Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: In Vitro Embryo Production Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Supplementation with EPS ID1 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Embryo Vitrification and Warming Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Children: Item: Title: Vitrification Protocol Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Warming Protocol Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Immunostaining for Differential Cell Count and DNA Fragmentation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Gene Expression Analysis by Reverse Transcription and Quantitative PCR (rt-qPCR) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Info: Title: Exopolysaccharide ID1 Improves Post-Warming Outcomes after Vitrification of In Vitro-Produced Bovine Embryos Author: Erika Alina Ordóñez-León, Iris Martínez-Rodero, Tania García-Martínez, Manel López-Béjar, Marc Yeste, Elena Mercade and Teresa Mogas Subject: This study aimed to assess the cryoprotectant role of exopolysaccharide (EPS) ID1, produced by Antarctic Pseudomonas sp., in the vitrification of in vitro-produced (IVP) bovine embryos. IVP day 7 (D7) and day 8 (D8) expanded blastocysts derived from cow or calf oocytes were vitrified without supplementation (EPS0) or supplemented with 10 g/mL (EPS10) or 100 g/mL (EPS100) EPS ID1. The effect of EPS ID1 was assessed in post-warming re-expansion and hatching rates, differential cell count, apoptosis rate, and gene expression. EPS100 re-expansion rates were significantly higher than those observed for the EPS0 and EPS10 treatments, regardless of culture length or oocyte source. EPS100 hatching rate was similar to the one of the fresh blastocysts except for those D7 blastocysts derived from calf oocytes. No differences were observed among EPS ID1 treatments when the inner cell mass, trophectoderm, and total cell number were assessed. Although apoptosis rates were higher (p 0.05) in vitrified groups compared to fresh embryos, EPS100 blastocysts had a lower number (p 0.05) of apoptotic nuclei than the EPS0 or EPS10 groups. No differences in the expression of BCL2, AQP3, CX43, and SOD1 genes between treatments were observed. Vitrification without EPS ID1 supplementation produced blastocysts with significantly higher BAX gene expression, whereas treatment with 100 g/mL EPS ID1 returned BAX levels to those observed in non-vitrified blastocysts. Our results suggest that 100 g/mL EPS ID1 added to the vitrification media is beneficial for embryo cryopreservation because it results in higher re-expansion and hatching ability and it positively modulates apoptosis. 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