Development of an Improved in vitro Model of Bovine Trophectoderm Differentiation

• DOI: 10.3389/fanim.2022.898808
• Publication Date: 2022-06-03T10:08:50Z
• Authors: M. Sofia Ortega; Jason A. Rizo; Jessica N. Drum; Eleanore V. O'Neil; Ky G. Pohler; Karl Kerns; Amanda Schmelze; Jonathan Green; Thomas E. Spencer
• Keywords: Conceptus; Placentation; Inner cell mass

The mechanisms regulating early stages of placentation and trophectoderm differentiation in the ruminant conceptus remain poorly understood. Here we present a model (TE) vitro from outgrowths individual derived embryos. Cell expressed markers mononucleate (MNC) binucleate (BNC) TE cells. percentage BNC ranged 14 to 39% as determined by flow cytometry. Pregnancy-associated glycoproteins (PAGs), produced BNC, were measured culture media on days 35 54. Continuous secretion PAGs was observed indicative functionality. Gene expression evaluated 20 embryo cell two different sires. Expression HAND1 , which is involved differentiation, CSH2 BNC-specific gene, altered between sires tested. Single-cell RNA-seq analysis day 40 revealed 11 distinct populations, with specific clusters genes lineage specification, proliferation, differentiation. In addition, whole -RNAseq performed confirmed sustained such some PAGs. developed bovine outgrowth found evidence for MNC continuous production PAGs, recapitulating key features placenta development. This can be used understand developmental biology cells, provide insights into paternal influences impact our understanding pregnancy loss cattle.