Generation of Hepatic Tissue Structures Using Multicellular Spheroid Culture

  • Fumiya Tao
  • Hirotaka Mihara
  • Nobuhiko KojimaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1905)


The hepatic functions of the hepatocytes in multicellular spheroid (MCS) are lower than those in the liver. One of the causes is that conventional hepatic MCSs do not reproduce liver-specific microstructures such as hepatic cord. It is necessary to design the inner structure of hepatic MCSs mimicking a structural feature of hepatic cord to represent further hepatic functions. Here we introduce a unique method to engineer the microarchitectures in the MCSs by formation of void spaces or filling of extracellular matrices (ECMs).

Key words

Multicellular spheroid Hepatocyte Methylcellulose Microstructure Alginate hydrogel bead Extracellular matrix 



This work was supported by JSPS KAKENHI under Grant Number 24710142 and AMED under Grant Number JP17be034302.


  1. 1.
    Laschke MW, Menger MD (2017) Life is 3D: boosting spheroid function for tissue engineering. Trends Biotechnol 35:133–144CrossRefGoogle Scholar
  2. 2.
    Kojima N, Takeuchi S, Sakai Y (2012) Rapid aggregation of heterogeneous cells and multiple-sized microspheres in methylcellulose medium. Biomaterials 33:4508–4514CrossRefPubMedGoogle Scholar
  3. 3.
    Kojima N, Takeuchi S, Sakai Y (2014) Engineering of pseudoislets: effect on insulin secretion activity by cell number, cell population, and microchannel networks. Transplant Proc 46:1161–1165CrossRefPubMedGoogle Scholar
  4. 4.
    Kojima N, Takeuchi S, Sakai Y (2014) Fabrication of microchannel networks in multicellular spheroids. Sensor Actuat B Chem 198:249–254CrossRefGoogle Scholar
  5. 5.
    Motoyama W, Sayo K, Mihara H et al (2016) Induction of hepatic tissues in multicellular spheroids composed of murine fetal hepatic cells and embedded hydrogel beads. Regen Ther 3:7–10CrossRefGoogle Scholar
  6. 6.
    Kamitori S, Ozeki Y, Kojima N (2016) β-Galactoside-mediated tissue organization during islet reconstitution. Regen Ther 3:11–14CrossRefGoogle Scholar
  7. 7.
    Sayo K, Aoki S, Kojima N (2016) Fabrication of bone marrow-like tissue in vitro from dispersed-state bone marrow cells. Regen Ther 3:32–37CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Life and Environmental System Science, Graduate School of NanobioscienceYokohama City UniversityYokohamaJapan

Personalised recommendations