Preparation of mechanically patterned hydrogels for controlling the self-condensation of cells

Takahisa Matsuzaki; Yuma Kawano; Momoka Horikiri; Yuko Shimokawa; Takashi Yamazaki; Nao Okuma; Hiroyuki Koike; Masaki Kimura; 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.xpro.2023.102471

Preparation of mechanically patterned hydrogels for controlling the self-condensation of cells

STAR Protoc. 2023 Sep 15;4(3):102471. doi: 10.1016/j.xpro.2023.102471. Epub 2023 Jul 28.

Authors

Takahisa Matsuzaki¹, Yuma Kawano², Momoka Horikiri³, Yuko Shimokawa², Takashi Yamazaki², Nao Okuma², Hiroyuki Koike⁴, Masaki Kimura⁴, Ryuzo Kawamura², Yosuke Yoneyama⁵, Yasuro Furuichi⁶, Fumihiko Hakuno⁷, Shin-Ichiro Takahashi⁷, Seiichiro Nakabayashi⁸, Satoshi Okamoto⁹, Hiromitsu Nakauchi¹⁰, Hideki Taniguchi⁹, Takanori Takebe¹¹, Hiroshi Y Yoshikawa¹²

Affiliations

¹ Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, Japan; Center for Future Innovation, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: matsuzaki@ap.eng.osaka-u.ac.jps.
² Department of Chemistry, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama 338-8570, Japan.
³ Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, Japan.
⁴ Department of Pediatrics, University of Cincinnati College of Medicine, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA; Division of Gastroenterology, Hepatology & 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.
⁵ Institute of Research, Division of Advanced Multidisciplinary Research, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
⁶ Department of Health Promotion Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo, Japan.
⁷ 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.
⁸ Department of Chemistry, Saitama University, Shimo-okubo 255, Sakura-ku, Saitama 338-8570, Japan; Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo 255, Sakura-ku, Saitama 338-8570, Japan.
⁹ Division of Regenerative Medicine, University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639, Japan; Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanazawa-ku 3-9, Yokohama, Kanagawa 236-0004, Japan.
¹⁰ Institute for Stem Cell Biology and Regenerative Medicine, School of Medicine, Stanford University, Stanford, CA 94305, USA; Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo, Japan.
¹¹ Department of Pediatrics, University of Cincinnati College of Medicine, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA; Division of Gastroenterology, Hepatology & 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; Institute of Research, Division of Advanced Multidisciplinary Research, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Premium Research Institute for Human Metaverse Medicine (WPI-PRIMe), and Division of Stem Cell and Organoid Medicine, Osaka University, Suita, Osaka 565-0871, Japan. Electronic address: takanori.takebe@cchmc.org.
¹² Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: hiroshi@ap.eng.osaka-u.ac.jp.

Abstract

Synthetic protocols providing mechanical patterns to culture substrate are essential to control the self-condensation of cells for organoid engineering. Here, we present a protocol for preparing hydrogels with mechanical patterns. We describe steps for hydrogel synthesis, mechanical evaluation of the substrate, and time-lapse imaging of cell self-organization. This protocol will facilitate the rational design of culture substrates with mechanical patterns for the engineering of various functional organoids. For complete details on the use and execution of this protocol, please refer to Takebe et al. (2015) and Matsuzaki et al. (2014, 2022).¹^,²^,³.

Keywords: Atomic Force Microscopy (AFM); Biophysics; Biotechnology and Bioengineering; Health Sciences; Material Sciences; Organoids; Stem Cells; Tissue Engineering.

MeSH terms

Hydrogels*
Organoids*

Substances

Hydrogels