Controlled Cytoplast Arrest and Morula Aggregation Enhance Development, Cryoresilience, and In Vivo Survival of Cloned Sheep Embryos

Simple item page
dc.citation.issue1
dc.citation.volume23
dc.contributor.authorMcLean ZL
dc.contributor.authorAppleby SJ
dc.contributor.authorFermin LM
dc.contributor.authorHenderson HV
dc.contributor.authorWei J
dc.contributor.authorWells DN
dc.contributor.authorOback B
dc.coverage.spatialUnited States
dc.date.accessioned2024-01-22T23:53:48Z
dc.date.accessioned2024-07-25T06:43:23Z
dc.date.available2021-02-02
dc.date.available2024-01-22T23:53:48Z
dc.date.available2024-07-25T06:43:23Z
dc.date.issued2021-02-02
dc.description.abstractZona-free somatic cell transfer (SCT) and embryo aggregation increase throughput and efficiency of cloned embryo and offspring production, respectively, but both approaches have not been widely adopted. Cloning efficiency is further improved by cell cycle coordination between the interphase donor cell and metaphase-arrested recipient cytoplast. This commonly involves inclusion of caffeine and omission of calcium to maintain high mitotic cyclin-dependent kinase activity and low calcium levels, respectively, in the nonactivated cytoplast. The aim of our study was to integrate these various methodological improvements into a single work stream that increases sheep cloning success. We show that omitting calcium during zona-free SCT improved blastocyst development from 6% to 13%, while caffeine treatment reduced spontaneous oocyte activation from 17% to 8%. In a retrospective analysis, morula aggregation produced high morphological quality blastocysts with better in vivo survival to term than nonaggregated controls (15% vs. 9%), particularly after vitrification (14% vs. 0%). By combining cytoplast cell cycle control with zona-free embryo reconstruction and aggregation, this novel SCT protocol maximizes the benefits of vitrification by producing more cryoresilient blastocysts. The presented cloning methodology is relatively easy to operate and further increases throughput and efficiency of cloned embryo and offspring production. Integration of additional reprogramming steps or alternate donor cells is straightforward, providing a flexible workflow that can be adapted to changing experimental requirements.
dc.description.confidentialfalse
dc.edition.editionFebruary 2021
dc.format.pagination14-25
dc.identifier.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/33529123
dc.identifier.citationMcLean ZL, Appleby SJ, Fermin LM, Henderson HV, Wei J, Wells DN, Oback B. (2021). Controlled Cytoplast Arrest and Morula Aggregation Enhance Development, Cryoresilience, and In Vivo Survival of Cloned Sheep Embryos.. Cell Reprogram. 23. 1. (pp. 14-25).
dc.identifier.doi10.1089/cell.2020.0078
dc.identifier.eissn2152-4998
dc.identifier.elements-typejournal-article
dc.identifier.issn2152-4971
dc.identifier.urihttps://mro.massey.ac.nz/handle/10179/70748
dc.languageeng
dc.publisherMary Ann Liebert, Inc
dc.publisher.urihttps://www.liebertpub.com/doi/10.1089/cell.2020.0078
dc.relation.isPartOfCell Reprogram
dc.rights(c) The author/sen
dc.rights.licenseCC BY-NCen
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectactivation
dc.subjectaggregation
dc.subjectcloning
dc.subjectcryopreservation
dc.subjectsheep
dc.subjectsomatic cell transfer
dc.subjectAnimals
dc.subjectBlastocyst
dc.subjectCloning, Organism
dc.subjectEmbryo, Mammalian
dc.subjectEmbryonic Development
dc.subjectMorula
dc.subjectNuclear Transfer Techniques
dc.subjectOocytes
dc.subjectSheep
dc.subjectVitrification
dc.titleControlled Cytoplast Arrest and Morula Aggregation Enhance Development, Cryoresilience, and In Vivo Survival of Cloned Sheep Embryos
dc.typeJournal article
pubs.elements-id444114
pubs.organisational-groupOther
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