Microfluidic confinement enhances phenotype and function of hepatocyte spheroids

Jong Hoon Choi, Lorena Loarca, Jose M. de Hoyos-Vega, Neda Dadgar, Kevin Loutherback, Vijay H. Shah, Gulnaz Stybayeva, Alexander Revzin

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

A number of cell culture approaches have been described for maintenance of primary hepatocytes. Forming hepatocytes into three-dimensional (3-D) spheroids is one well-accepted method for extending epithelial phenotype of these cells. Our laboratory has previously observed enhanced function of two-dimensional (2-D, monolayer) hepatocyte cultures in microfluidic devices due to increased production of several hepato-inductive growth factors, including hepatocyte growth factor (HGF). In the present study, we wanted to test a hypothesis that culturing hepatocyte spheroids (3-D) in microfluidic devices will also result in enhanced phenotype and function. To test this hypothesis, we fabricated devices with small and large volumes. Both types of devices included a microstructured floor containing arrays of pyramidal wells to promote assembly of hepatocytes into spheroids with individual diameters of ~100 μm. The hepatocyte spheroids were found to be more functional, as evidenced by higher level of albumin synthesis, bile acid production, and hepatic enzyme expression, in low-volume compared with large-volume devices. Importantly, high functionality of spheroid cultures correlated with elevated levels of HGF secretion. Although decay of hepatic function (albumin secretion) was observed over the course 3 wk, this behavior could be abrogated by inhibiting TGF- β1 signaling. With TGF- β1 inhibitor, microfluidic hepatocyte spheroid cultures maintained high and stable levels of albumin synthesis over the course of 4 wk. To further highlight utility of this culture platform for liver disease modeling, we carried out alcohol injury experiments in microfluidic devices and tested protective effects of interleukin-22: a potential therapy for alcoholic hepatitis.

Original languageEnglish (US)
Pages (from-to)C552-C560
JournalAmerican Journal of Physiology - Cell Physiology
Volume319
Issue number3
DOIs
StatePublished - Sep 2020

Keywords

  • Alcohol injury
  • Hepatocyte spheroids
  • Microfluidic cultures

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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