Mechanical guidance of self-condensation patterns of differentiating progeny

Takahisa Matsuzaki; Yuko Shimokawa; Hiroyuki Koike; Masaki Kimura; Yuma Kawano; Nao Okuma; Ryuzo Kawamura; Yosuke Yoneyama; Yasuro Furuichi; Fumihiko Hakuno; Shin-Ichiro Takahashi; Seiichiro Nakabayashi; Satoshi Okamoto; Hiromitsu Nakauchi; Hideki Taniguchi; Takanori Takebe; Hiroshi Y Yoshikawa

doi:10.1016/j.isci.2022.105109

Mechanical guidance of self-condensation patterns of differentiating progeny

iScience. 2022 Sep 27;25(10):105109. doi: 10.1016/j.isci.2022.105109. eCollection 2022 Oct 21.

Authors

Takahisa Matsuzaki^{1

2

3

4}, Yuko Shimokawa⁴, Hiroyuki Koike^{5

6}, Masaki Kimura^{5

6}, Yuma Kawano⁴, Nao Okuma⁴, Ryuzo Kawamura⁴, Yosuke Yoneyama^{7

8}, Yasuro Furuichi⁹, Fumihiko Hakuno⁸, Shin-Ichiro Takahashi⁸, Seiichiro Nakabayashi^{3

4}, Satoshi Okamoto^{10

11}, Hiromitsu Nakauchi^{12

13}, Hideki Taniguchi^{10

11}, Takanori Takebe^{5

6

7}, Hiroshi Y Yoshikawa^{2

3

4}

Affiliations

¹ Center for Future Innovation, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, Japan.
² Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, Japan.
³ Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo 255, Sakura-Ku, Saitama 338-8570, Japan.
⁴ Department of Chemistry, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama 338-8570, Japan.
⁵ Department of Pediatrics, University of Cincinnati College of Medicine, 3333 Burnet Avenue, Cincinnati, OH 45229- 3039, USA.
⁶ Division of Gastroenterology, Hepatology and Nutrition, Developmental Biology, Center for Stem Cell and Organoid Medicine (CuSTOM), Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA.
⁷ Institute of Research, Division of Advanced Multidisciplinary Research, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-Ku, Tokyo 113-8510, Japan.
⁸ Departments of Animal Sciences and Applied Biological Chemistry, Graduate School of Agriculture and Life Sciences, the University of Tokyo, Bunkyo-Ku, Tokyo 113-8657, Japan.
⁹ Department of Health Promotion Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo, Japan.
¹⁰ Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanazawa-Ku 3-9, Yokohama, Kanagawa, 236-0004, Japan.
¹¹ Division of Regenerative Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, University of Tokyo, 4-6-1, Shirokanedai, Minato-Ku, Tokyo 108-8639, Japan.
¹² Institute for Stem Cell Biology and Regenerative Medicine, School of Medicine, Stanford University, Stanford, CA 94305, USA.
¹³ Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo, Minato-Ku, Tokyo, Japan.

Abstract

Spatially controlled self-organization represents a major challenge for organoid engineering. We have developed a mechanically patterned hydrogel for controlling self-condensation process to generate multi-cellular organoids. We first found that local stiffening with intrinsic mechanical gradient (IG > 0.008) induced single condensates of mesenchymal myoblasts, whereas the local softening led to stochastic aggregation. Besides, we revealed the cellular mechanism of two-step self-condensation: (1) cellular adhesion and migration at the mechanical boundary and (2) cell-cell contraction driven by intercellular actin-myosin networks. Finally, human pluripotent stem cell-derived hepatic progenitors with mesenchymal/endothelial cells (i.e., liver bud organoids) experienced collective migration toward locally stiffened regions generating condensates of the concave to spherical shapes. The underlying mechanism can be explained by force competition of cell-cell and cell-hydrogel biomechanical interactions between stiff and soft regions. These insights will facilitate the rational design of culture substrates inducing symmetry breaking in self-condensation of differentiating progeny toward future organoid engineering.

Keywords: Biomaterials; Biomechanical engineering; Mechanobiology.