Secreted metabolome of porcine blastocysts encapsulated within

Abstract

Context. The exact mechanisms regulating the initiation of porcine conceptus elongation are not known due to the complexity of the uterine environment. Aims. To identify contributing factors for initiation of conceptus elongation in vitro, this study evaluated differential metabolite abundance within media following culture of blastocysts within unmodified alginate (ALG) or Arg-Gly-Asp (RGD)-modified alginate hydrogel culture systems. Methods. Blastocysts were harvested from pregnant gilts, encapsulated within ALG or RGD or as non-encapsulated control blastocysts (CONT), and cultured. At the termination of 96 h culture, media were separated into blastocyst media groups: non-encapsulated control blastocysts (CONT); ALG and RGD blastocysts with no morphological change (ALG− and RGD−); ALG and RGD blastocysts with morphological changes (ALG+ and RGD+) and evaluated for non-targeted metabolomic profiling by liquid chromatography (LC)-mass spectrometry (MS) techniques and gas chromatography-(GC-MS). Key results. Analysis of variance identified 280 (LC-MS) and 1 (GC-MS) compounds that differed (P < 0.05), of which 134 (LC-MS) and 1 (GC-MS) were annotated. Metabolites abundance between ALG+ vs ALG−, RGD+ vs RGD−, and RGD+ vs ALG+ were further investigated to identify potential differences in metabolic processes during the initiation of elongation. Conclusions. This study identified changes in phospholipid, glycosphingolipid, lipid signalling, and amino acid metabolic processes as potential RGD-independent mechanisms of elongation and identified changes in lysophosphatidylcholine and sphingolipid secretions during RGD-mediated elongation. Implications. These results illustrate changes in phospholipid and sphingolipid metabolic processes and secretions may act as mediators of the RGD-integrin adhesion that promotes porcine conceptus elongation.

Understanding bovine embryo elongation: a transcriptomic study of trophoblastic vesicles

“…Such data could be combined with other models of tissue elongation in order to highlight common themes, including patterning, spatiotemporal cell fate, or physical forces that enable proper morphogenesis ( Horne-Badovinac, 2014 ; Weng et al, 2022 ; Adler et al, 2023 ). Taken together, our results provide useful information for the future analyses of the factors that coordinate the conserved morphogenetic processes at work during bovine conceptus development, processes that one may evaluate in the coming years through novel cell culture modes ( Walsh et al, 2023 ), cocultures ( Wei et al, 2023 ), or in silico predictive models ( Butt et al, 2023 ).…”

Opportunities involving microfluidics and 3D culture systems to the in vitro embryo production

“…The same research group conjugated alginate hydrogels with Arg-Gly-Asp and it increased the embryo survival and induced morphological changes, when compared to the normal alginate hydrogel ( Laughlin et al, 2017 ). This model was recently used to study metabolome changes during the elongation process, which identified changes in secreted metabolites that may be correlated with the elongation mechanism ( Walsh et al, 2023 ).…”

“…In (C) bovine embryos were cultured in groups of 5 encapsulated inside an alginate bead or in conventional culture(Yániz et al, 2002). In (D) porcine blastocysts were cultured in a two-layered alginate bead to support its development after hatching(Sargus-Patino et al, 2014;Laughlin et al, 2017;Walsh et al, 2023). In (E) bovine blastocysts in the hatching stage were cultured in an alginate bead or alginate overlay(Zhao et al, 2015).Figure made with Biorender.…”