Generation of organized germ layers from a single mouse embryonic stem cell

Yeh-Chuin Poh, Junwei Chen, Ying Hong, Haiying Yi, Shuang Zhang, Junjian Chen, Douglas C. Wu, Lili Wang, Qiong Jia, Rishi Singh, Wenting Yao, Youhua Tan, Arash Tajik, Tetsuya S. Tanaka and Ning Wang ()
Additional contact information
Yeh-Chuin Poh: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Junwei Chen: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Ying Hong: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Haiying Yi: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Shuang Zhang: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Junjian Chen: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Douglas C. Wu: College of Engineering, University of Illinois at Urbana-Champaign
Lili Wang: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Qiong Jia: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Rishi Singh: College of Engineering, University of Illinois at Urbana-Champaign
Wenting Yao: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Youhua Tan: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology
Arash Tajik: College of Engineering, University of Illinois at Urbana-Champaign
Tetsuya S. Tanaka: University of Notre Dame
Ning Wang: Laboratory for Cell Biomechanics and Regenerative Medicine, School of Life Science and Technology, Huazhong University of Science and Technology

Nature Communications, 2014, vol. 5, issue 1, 1-12

Abstract: Abstract Mammalian inner cell mass cells undergo lineage-specific differentiation into germ layers of endoderm, mesoderm and ectoderm during gastrulation. It has been a long-standing challenge in developmental biology to replicate these organized germ layer patterns in culture. Here we present a method of generating organized germ layers from a single mouse embryonic stem cell cultured in a soft fibrin matrix. Spatial organization of germ layers is regulated by cortical tension of the colony, matrix dimensionality and softness, and cell–cell adhesion. Remarkably, anchorage of the embryoid colony from the 3D matrix to collagen-1-coated 2D substrates of ~1 kPa results in self-organization of all three germ layers: ectoderm on the outside layer, mesoderm in the middle and endoderm at the centre of the colony, reminiscent of generalized gastrulating chordate embryos. These results suggest that mechanical forces via cell–matrix and cell–cell interactions are crucial in spatial organization of germ layers during mammalian gastrulation. This new in vitro method could be used to gain insights on the mechanisms responsible for the regulation of germ layer formation.

Date: 2014
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DOI: 10.1038/ncomms5000

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