{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:27:15Z","timestamp":1785414435772,"version":"3.56.0"},"reference-count":44,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T00:00:00Z","timestamp":1778803200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ANID-Chile","award":["11230091"],"award-info":[{"award-number":["11230091"]}]},{"award":["11230091"],"award-info":[{"award-number":["11230091"]}],"id":[{"id":"https:\/\/ror.org\/02ap3w078","id-type":"ROR","asserted-by":"publisher"}]},{"name":"ANID\u2013FONDECYT","award":["1250823"],"award-info":[{"award-number":["1250823"]}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"crossref","award":["FPP24-0006"],"award-info":[{"award-number":["FPP24-0006"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"crossref"}]},{"award":["FPP24-0006"],"award-info":[{"award-number":["FPP24-0006"]}],"id":[{"id":"https:\/\/ror.org\/02ddkpn78","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Haploid embryos constitute a valuable model for genetic and epigenetic studies; however, their developmental competence is reduced compared with diploid counterparts. This study evaluated whether supplementation of the culture medium with specific small molecules could improve developmental competence and outgrowth establishment of parthenogenetic haploid embryos. The effects of TGF-\u03b2 inhibition (A83-01), WNT pathway modulation (CHIR99021 and IWR-1), and activin A (AA) supplementation were assessed from the morula stage onward under serum-free conditions. A83-01 treatment did not improve blastocyst formation or morphology and was associated with reduced total cell numbers relative to IVF controls. CHIR99021 supplementation increased the number of SOX2-positive cells compared with IWR-1 and vehicle-treated embryos, suggesting partial support of pluripotency; however, overall developmental progression remained inferior to diploid controls. In contrast, activin A significantly increased the proportion of haploid morulae developing into blastocysts and improved hatching rates. Nevertheless, AA supplementation did not restore CDX2-positive cell numbers or total cell counts to diploid levels. Furthermore, neither CHIR99021 nor AA affect DNA fragmentation levels, although a tendency toward increased TUNEL-positive cells was observed. Activin A treatment also failed to improve embryonic outgrowth formation. Collectively, these findings demonstrate that although activin A enhances blastocyst yield and hatching in bovine haploid embryos, modulation of TGF-\u03b2 or WNT signaling alone is insufficient to restore diploid-like proliferative developmental competence.<\/jats:p>","DOI":"10.3390\/ani16101517","type":"journal-article","created":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T14:19:29Z","timestamp":1778854769000},"page":"1517","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effect of Small Molecules on Blastocyst Development and Outgrowth Establishment of Bovine Haploid Parthenogenetic Embryos"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9713-4248","authenticated-orcid":false,"given":"Luis","family":"Aguila","sequence":"first","affiliation":[{"name":"Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811322, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2591-1544","authenticated-orcid":false,"given":"Rodrigo","family":"Castillo","sequence":"additional","affiliation":[{"name":"Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811322, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-4801-933X","authenticated-orcid":false,"given":"Felipe","family":"P\u00e9rez-Garc\u00eda","sequence":"additional","affiliation":[{"name":"Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811322, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0851-8608","authenticated-orcid":false,"given":"Favian","family":"Treulen","sequence":"additional","affiliation":[{"name":"Escuela de Tecnolog\u00eda M\u00e9dica, Facultad de Medicina y Ciencias de la Salud, Universidad Mayor, Temuco 4801043, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-7190-3927","authenticated-orcid":false,"given":"Cecilia","family":"Valencia","sequence":"additional","affiliation":[{"name":"Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811322, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2009-5863","authenticated-orcid":false,"given":"Felipe","family":"Perecin","sequence":"additional","affiliation":[{"name":"Departamento de Medicina Veterin\u00e1ria, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de S\u00e3o Paulo, Pirassununga 13635-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8766-0148","authenticated-orcid":false,"given":"Lawrence Charles","family":"Smith","sequence":"additional","affiliation":[{"name":"Centre de Recherche en Reproduction et F\u00e9rtilit\u00e9 (CRRF), Universit\u00e9 de Montr\u00e9al, Saint-Hyacinthe, QC J2S 2M2, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5442-1693","authenticated-orcid":false,"given":"Maria Elena","family":"Arias","sequence":"additional","affiliation":[{"name":"Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811322, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ricardo","family":"Felmer","sequence":"additional","affiliation":[{"name":"Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agriculture and Environmental Sciences, Universidad de La Frontera, Temuco 4811322, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1071\/RD08024","article-title":"Putative imprinted gene expression in uniparental bovine embryo models","volume":"20","author":"Cruz","year":"2008","journal-title":"Reprod. 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