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Probing morphological,genetic and metabolomic changes of in vitro embryo development in a microfluidic device |
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| Authors: | Vanessa Mancini Paul J McKeegan Alexandra C Schrimpe-Rutledge Simona G Codreanu Stacy D Sherrod John A McLean Helen M Picton Virginia Pensabene |
| Institution: | 1. School of Electronic and Electrical Engineering, University of Leeds, Leeds, UK
Contribution: Data curation, Formal analysis, ?Investigation, Validation, Writing - review & editing;2. Reproduction and Early Development Research Group, Discovery and Translational Science Department, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, UK Centre for Anatomical and Human Sciences, Hull York Medical School, University of Hull, Hull, UK Contribution: Data curation, Formal analysis, ?Investigation, Methodology, Validation, Visualization;3. Center for Innovative Technology (CIT), Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA Contribution: Data curation, Methodology, Validation, Visualization;4. Center for Innovative Technology (CIT), Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA Contribution: Conceptualization, Data curation, Formal analysis, ?Investigation, Methodology, Visualization;5. Center for Innovative Technology (CIT), Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA Contribution: Conceptualization, Methodology, Supervision, Writing - review & editing;6. Center for Innovative Technology (CIT), Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA Contribution: Conceptualization, Funding acquisition, Methodology, Resources, Writing - review & editing;7. Reproduction and Early Development Research Group, Discovery and Translational Science Department, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, UK Contribution: Conceptualization, Data curation, Funding acquisition, Methodology, Resources, Supervision, Writing - review & editing;8. School of Electronic and Electrical Engineering, University of Leeds, Leeds, UK |
| Abstract: | Assisted reproduction technologies for clinical and research purposes rely on a brief in vitro embryo culture which, despite decades of progress, remain suboptimal in comparison to the physiological environment. One promising tool to improve this technique is the development of bespoke microfluidic chambers. Here we present and validate a new microfluidic device in polydimethylsiloxane (PDMS) for the culture of early mouse embryos. Device material and design resulted embryo compatible and elicit minimal stress. Blastocyst formation, hatching, attachment and outgrowth formation on fibronectin-coated devices were similar to traditional microdrop methods. Total blastocyst cell number and allocation to the trophectoderm and inner cell mass lineages were unaffected. The devices were designed for culture of 10–12 embryos. Development rates, mitochondrial polarization and metabolic turnover of key energy substrates glucose, pyruvate and lactate were consistent with groups of 10 embryos in microdrop controls. Increasing group size to 40 embryos per device was associated with increased variation in development rates and altered metabolism. Device culture did not perturb blastocyst gene expression but did elicit changes in embryo metabolome, which can be ascribed to substrate leaching from PDMS and warrant further investigation. |
| Keywords: | assisted reproductive technologies embryo culture IVF mice microfluidics |