The method of mouse embryoid body establishment affects structure and developmental gene expression

Tissue & Cell
A MogiK Sasaki

Abstract

To investigate formation of the three primary germ layers in mouse embryoid bodies (EBs), we observed changes in structure and gene expression over a 7-day culture period. We compared these changes using two methods for EB formation: hanging drop (HD) and static suspension culture (SSC). Light microscopy showed that a stratified columnar epithelial layer developed on the surface of EBs formed using the HD method. From Day 3 in culture, ultrastructural changes occurred in the aligned cellular membranes. Condensation of actin filaments was followed by formation of complicated adherent junctions and dilatation of intercellular canaliculi containing well-developed microvilli. These changes were more marked in EBs formed by the HD method than the SSC method. On Day 5 of culture, Brachyury gene expression, a marker for mesoderm formation, was detected only with the HD method. Nestin, an ectoderm marker, and Foxa2, an endoderm marker, were expressed with both methods. These results suggest that in EBs formed with the HD method, actin formation and Brachyury gene expression mark the transition from two to three primary germ layers. Additionally, the HD method promotes more rapid and complete development of mouse EBs than does the SSC m...Continue Reading

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Citations

Mar 29, 2011·Gene Expression Patterns : GEP·Rashmi TripathiAnton J Enright
Apr 3, 2009·Biologicals : Journal of the International Association of Biological Standardization·J QinZ M Cai
Oct 10, 2009·Biotechnology and Bioengineering·Carolyn Y SargentTodd C McDevitt
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Jun 3, 2016·Cellular Reprogramming·Yasumasa ShirouzuShoichiro Sumi
Sep 16, 2016·PloS One·Daihachiro TomotsuneKatsunori Sasaki
Nov 12, 2009·Lab on a Chip·Diep NguyenMichelle Khine
Nov 6, 2014·BioResearch Open Access·Anne-Marie CourtotAnnelise Bennaceur-Griscelli
Sep 16, 2009·Tissue Engineering. Part C, Methods·David GothardLee D Buttery
Nov 23, 2010·Tissue Engineering. Part a·Limor Zwi-DantsisLior Gepstein

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