Reconstruction of Hepatic Tissue Structures Using Interstitial Flow in a Microfluidic Device

  • Ryo SudoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1905)


Construction of three-dimensional (3D) hepatic tissue structures is important for in vitro tissue engineering of the liver, because 3D culture of hepatocytes is critical for the maintenance of liver-specific functions. Although conventional 3D culture methods are useful for constructing 3D hepatic tissue structures, the precise control of culture microenvironments is required to construct more physiological tissues in vitro. Recent advances in microfluidics technologies have allowed us to utilize microfluidic devices for hepatic cell culture, which opened the door for creating more physiological 3D culture models of the liver. Here, we describe the method for the construction of hepatic tissue structures using a microfluidic device which has a 3D gel region with adjacent microchannels. Primary rat hepatocytes are seeded into a microchannel in a microfluidic device. The cells are then cultured in interstitial flow conditions, which leads to the construction of 3D tissue structures.

Key words

Microfluidic device Interstitial flow 3D culture 



This work was supported by JSPS KAKENHI Grant Number 16H03173 to R.S.


  1. 1.
    Sudo R (2014) Multiscale tissue engineering for liver reconstruction. Organogenesis 10:216–224CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Chung S, Sudo R, Vickerman V, Zervantonakis IK, Kamm RD (2010) Microfluidic platforms for studies of angiogenesis, cell migration, and cell-cell interactions. Ann Biomed Eng 38:1164–1177CrossRefGoogle Scholar
  3. 3.
    Zervantonakis IK, Kothapalli CR, Chung S, Sudo R, Kamm RD (2011) Microfluidic devices for studying heterotypic cell-cell interactions and tissue specimen cultures under controlled microenvironments. Biomicrofluidics 5:13406CrossRefPubMedGoogle Scholar
  4. 4.
    Shin Y, Han S, Jeon JS, Yamamoto K, Zervantonakis IK, Sudo R, Kamm RD, Chung S (2012) Microfluidic assay for simultaneous culture of multiple cell types on surfaces or within hydrogels. Nat Protoc 7:1247–1259CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Sudo R, Chung S, Zervantonakis IK, Vickerman V, Toshimitsu Y, Griffith LG, Kamm RD (2009) Transport-mediated angiogenesis in 3D epithelial coculture. FASEB J 23:2155–2164CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Yamamura N, Sudo R, Ikeda M, Tanishita K (2007) Effects of the mechanical properties of collagen gel on the in vitro formation of microvessel networks by endothelial cells. Tissue Eng 13:1443–1453CrossRefPubMedGoogle Scholar
  7. 7.
    Chung S, Sudo R, Mack PJ, Wan CR, Vickerman V, Kamm RD (2009) Cell migration into scaffolds under co-culture conditions in a microfluidic platform. Lab Chip 9:269–275CrossRefPubMedGoogle Scholar
  8. 8.
    Chung S, Sudo R, Zervantonakis IK, Rimchala T, Kamm RD (2009) Surface treatment induced three dimensional capillary morphogenesis in a microfluidic platform. Adv Mater 21:4863–4867CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of System Design EngineeringKeio UniversityYokohamaJapan

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