Efficiency of embryo complementation and pluripotency maintenance following multiple passaging of in vitro-derived bovine embryos
Maura S. McGraw A , Jordan A. Bishman A and Bradford W. Daigneault
A *
A
Abstract
Current methods to obtain bovine embryos of high genetic merit include approaches that require skilled techniques for low-efficiency cloning strategies.
The overall goal herein was to identify the efficacy of alternative methods for producing multiple embryos through blastomere complementation while determining maintenance of cell pluripotency.
Bovine oocytes were fertilised in vitro to produce 4-cell embryos from which blastomeres were isolated and cultured as 2-cell aggregates using a well-of-the-well system. Aggregates were returned to incubation up to 7 days (Passage 1). A second passage of complement embryos was achieved by splitting 4-cell Passage 1 embryos. Passaged embryos reaching the blastocyst stage were characterised for cell number and cell lineage specification in replicate with non-reconstructed zona-intact embryos.
Passage 1 and 2 embryo complements yielded 29% and 25% blastocyst development, respectively. Passage 1 embryos formed blastocysts, but with a reduction in expression of SOX2 and decreased size compared to non-reconstructed zona-intact embryos. Passage 2 embryos had a complete lack of SOX2 expression and a reduction in transcript abundance of SOX2 and SOX17, suggesting loss of pluripotency markers that primarily affected inner cell mass (ICM) and hypoblast formation.
In vitro fertilised bovine embryos can be reconstructed with multiple passaging to generate genetically identical embryos. Increased passaging drives trophectoderm cell lineage specification while compromising ICM formation.
These results may provide an alternative strategy for producing genetically identical bovine embryos through blastomere complementation with applications towards the development of trophoblast and placental models of early development.
Keywords: aggregate, blastomere, bovine, clone, complementation, embryo, inner cell mass, pluripotent, trophectoderm.
References
Barak Y, Nelson MC, Ong ES, Jones YZ, Ruiz-Lozano P, Chien KR, Koder A, Evans RM (1999) PPARγ is required for placental, cardiac, and adipose tissue development. Molecular Cell 4, 585-595.
| Crossref | Google Scholar | PubMed |
Berg DK, Smith CS, Pearton DJ, Wells DN, Broadhurst R, Donnison M, Pfeffer PL (2011) Trophectoderm lineage determination in cattle. Developmental Cell 20, 244-255.
| Crossref | Google Scholar | PubMed |
Carreiro LE, Santos GSd, Luedke FE, Goissis MD (2021) Cell differentiation events in pre-implantation mouse and bovine embryos. Animal Reproduction 18, e20210054.
| Crossref | Google Scholar |
Chavatte-Palmer P, de Sousa N, Laigre P, Camous S, Ponter AA, Beckers J-F, Heyman Y (2006) Ultrasound fetal measurements and pregnancy associated glycoprotein secretion in early pregnancy in cattle recipients carrying somatic clones. Theriogenology 66, 829-840.
| Crossref | Google Scholar | PubMed |
Choe Y-H, Sorensen J, Garry DJ, Garry MG (2022) Blastocyst complementation and interspecies chimeras in gene edited pigs. Frontiers in Cell and Developmental Biology 10, 1065536.
| Crossref | Google Scholar |
Daigneault BW, Rajput S, Smith GW, Ross PJ (2018a) Embryonic POU5F1 is required for expanded bovine blastocyst formation. Scientific Reports 8, 7753.
| Crossref | Google Scholar | PubMed |
Daigneault BW, Vilarino M, Rajput SK, Frum T, Smith GW, Ross PJ (2018b) CRISPR editing validation, immunostaining and DNA sequencing of individual fixed bovine embryos. BioTechniques 65, 281-283.
| Crossref | Google Scholar | PubMed |
Farin PW, Piedrahita JA, Farin CE (2006) Errors in development of fetuses and placentas from in vitro-produced bovine embryos. Theriogenology 65, 178-191.
| Crossref | Google Scholar | PubMed |
Ferré LB, Kjelland ME, Strøbech LB, Hyttel P, Mermillod P, Ross PJ (2020) Review: Recent advances in bovine in vitro embryo production: reproductive biotechnology history and methods. Animal 14, 991-1004.
| Crossref | Google Scholar | PubMed |
Freedman BS (2018) Hopes and difficulties for blastocyst complementation. Nephron 138, 42-47.
| Crossref | Google Scholar | PubMed |
Gambini A, Jarazo J, Olivera R, Salamone DF (2012) Equine cloning: in vitro and in vivo development of aggregated embryos. Biology of Reproduction 87, 1-5.
| Crossref | Google Scholar |
Goissis MD, Cibelli JB (2014) Functional characterization of CDX2 during bovine preimplantation development in vitro. Molecular Reproduction and Development 81, 962-970.
| Crossref | Google Scholar | PubMed |
Golding MC (2012) Generation of trophoblast stem cells. In ‘Methods in molecular biology’. (Ed. N Engel) pp. 49–59. (Humana Press) doi:10.1007/978-1-62703-011-3_3
Halstead MM, Ma X, Zhou C, Schultz RM, Ross PJ (2020) Chromatin remodeling in bovine embryos indicates species-specific regulation of genome activation. Nature Communications 11, 4654.
| Crossref | Google Scholar | PubMed |
Hashimoto H, Eto T, Yamamoto M, Yagoto M, Goto M, Kagawa T, Kojima K, Kawai K, Akimoto T, Takahashi R-I (2019) Development of blastocyst complementation technology without contributions to gametes and the brain. Experimental Animals 68, 361-370.
| Crossref | Google Scholar | PubMed |
Hiriart MI, Bevacqua RJ, Canel NG, Fernández-Martín R, Salamone DF (2013) Production of chimeric embryos by aggregation of bovine egfp eight-cell stage blastomeres with two-cell fused and asynchronic embryos. Theriogenology 80, 357-364.
| Crossref | Google Scholar | PubMed |
Johnson W, Loskutoff N, Plante Y, Betteridge K (1995) Production of four identical calves by the separation of blastomeres from an in vitro derived four-cell embryo. Veterinary Record 137, 15-16.
| Crossref | Google Scholar | PubMed |
Kurosaka S, Eckardt S, Ealy AD, McLaughlin KJ (2007) Regulation of blastocyst stage gene expression and outgrowth interferon τ activity of somatic cell clone aggregates. Cloning and Stem Cells 9, 630-641.
| Crossref | Google Scholar | PubMed |
Lundin K, Bergh C, Hardarson T (2001) Early embryo cleavage is a strong indicator of embryo quality in human IVF. Human Reproduction 16, 2652-2657.
| Crossref | Google Scholar | PubMed |
Luo L, Shi Y, Wang H, Wang Z, Dang Y, Li S, Wang S, Zhang K (2022) Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency. PLoS Genetics 18, e1010307.
| Crossref | Google Scholar | PubMed |
McGraw MS, Rajput SK, Daigneault BW (2024) PPAR-gamma influences developmental competence and trophectoderm lineage specification in bovine embryos. Reproduction 167(2), e230334.
| Crossref | Google Scholar | PubMed |
Ménézo YJR, Hérubel F (2002) Mouse and bovine models for human IVF. Reproductive BioMedicine Online 4, 170-175.
| Crossref | Google Scholar | PubMed |
Reese ST, Franco GA, Poole RK, Hood R, Fernadez Montero L, Oliveira Filho RV, Cooke RF, Pohler KG (2020) Pregnancy loss in beef cattle: a meta-analysis. Animal Reproduction Science 212, 106251.
| Crossref | Google Scholar | PubMed |
Rossant J (2011) A mouse is not a cow. Nature 471, 457-458.
| Crossref | Google Scholar |
Shi Y, Hu B, Wang Z, Wu X, Luo L, Li S, Wang S, Zhang K, Wang H (2023) Functional role of GATA3 and CDX2 in lineage specification during bovine early embryonic development. Reproduction 165, 325-333.
| Crossref | Google Scholar |
Somfai T, Inaba Y, Aikawa Y, Ohtake M, Kobayashi S, Konishi K, Imai K (2010) Relationship between the length of cell cycles, cleavage pattern and developmental competence in bovine embryos generated by in vitro fertilization or parthenogenesis. Journal of Reproduction and Development 56, 200-207.
| Crossref | Google Scholar | PubMed |
Tagawa M, Matoba S, Narita M, Saito N, Nagai T, Imai K (2008) Production of monozygotic twin calves using the blastomere separation technique and Well of the Well culture system. Theriogenology 69, 574-582.
| Crossref | Google Scholar | PubMed |
Tríbulo P, Rivera RM, Ortega Obando MS, Jannaman EA, Hansen PJ (2019) Production and culture of the bovine embryo. In ‘Methods in molecular biology’. (Ed. J Herrick) pp. 115–129. doi:10.1007/978-1-4939-9566-0_8
Vajta G, Peura TT, Holm P, Paldi A, Greve T, Trounson AO, Callesen H (2000) New method for culture of zona-included or zona-free embryos: the well of the well (WOW) system. Molecular Reproduction and Development 55, 256-264.
| Crossref | Google Scholar | PubMed |
Vajta G, Korösi T, Du Y, Nakata K, Ieda S, Kuwayama M, Nagy ZP (2008) The well-of-the-well system: an efficient approach to improve embryo development. Reproductive BioMedicine Online 17, 73-81.
| Crossref | Google Scholar | PubMed |
Wang Y, Ming H, Yu L, Li J, Zhu L, Sun H-X, Pinzon-Arteaga CA, Wu J, Jiang Z (2023) Establishment of bovine trophoblast stem cells. Cell Reports 42, 112439.
| Crossref | Google Scholar | PubMed |
Warzych E, Pawlak P, Lechniak D, Madeja ZE (2020) WNT signalling supported by MEK/ERK inhibition is essential to maintain pluripotency in bovine preimplantation embryo. Developmental Biology 463, 63-76.
| Crossref | Google Scholar | PubMed |
Whitworth KM, Prather RS (2010) Somatic cell nuclear transfer efficiency: how can it be improved through nuclear remodeling and reprogramming? Molecular Reproduction and Development 77, 1001-1015.
| Crossref | Google Scholar | PubMed |
Wicklow E, Blij S, Frum T, Hirate Y, Lang RA, Sasaki H, Ralston A (2014) HIPPO pathway members restrict SOX2 to the inner cell mass where it promotes ICM fates in the mouse blastocyst. PLoS Genetics 10, e1004618.
| Crossref | Google Scholar | PubMed |
Willadsen SM (1979) A method for culture of micromanipulated sheep embryos and its use to produce monozygotic twins. Nature 277, 298-300.
| Crossref | Google Scholar | PubMed |
Willadsen S, Polge C (1981) Attempts to produce monozygotic quadruplets in cattle by blastomere separation. Veterinary Record 108, 211-213.
| Crossref | Google Scholar | PubMed |
Wu J, Platero-Luengo A, Sakurai M, Sugawara A, Gil MA, Yamauchi T, Suzuki K, Bogliotti YS, Cuello C, Morales Valencia M, Okumura D, Luo J, Vilariño M, Parrilla I, Soto DA, Martinez CA, Hishida T, Sánchez-Bautista S, Martinez-Martinez ML, Wang H, Nohalez A, Aizawa E, Martinez-Redondo P, Ocampo A, Reddy P, Roca J, Maga EA, Esteban CR, Berggren WT, Nuñez Delicado E, Lajara J, Guillen I, Guillen P, Campistol JM, Martinez EA, Ross PJ, Izpisua Belmonte JC (2017) Interspecies chimerism with mammalian pluripotent stem cells. Cell 168, 473-486.e15.
| Crossref | Google Scholar |
