Microscale collagen and fibroblast interactions enhance primary human hepatocyte functions in three-dimensional models

David A. Kukla, Alexandra L. Crampton, David K. Wood, Salman R. Khetani

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Human liver models that are three-dimensional (3D) in architecture are indispensable for compound metabolism/ toxicity screening, to model liver diseases for drug discovery, and for cell-based therapies; however, further development of such models is needed to maintain high levels of primary human hepatocyte (PHH) functions for weeks to months. Therefore, here we determined how microscale 3D collagen I presentation and fibroblast interaction affect the longevity of PHHs. High-throughput droplet microfluidics was utilized to generate reproducibly sized (~300-µm diameter) microtissues containing PHHs encapsulated in collagen I ± supportive fibroblasts, namely, 3T3-J2 murine embryonic fibroblasts or primary human hepatic stellate cells (HSCs); self-assembled spheroids and bulk collagen gels (macrogels) containing PHHs served as controls. Hepatic functions and gene expression were subsequently measured for up to 6 weeks. We found that microtissues placed within multiwell plates rescued PHH functions at 2- to 30-fold higher levels than spheroids or macrogels. Further coating of PHH microtissues with 3T3-J2s led to higher hepatic functions than when the two cell types were either coencapsulated together or when HSCs were used for the coating instead. Importantly, the 3T3-J2-coated PHH microtissues displayed 6+ weeks of relatively stable hepatic gene expression and function at levels similar to freshly thawed PHHs. Lastly, microtissues responded in a clinically relevant manner to drug-mediated cytochrome P450 induction or hepatotoxicity. In conclusion, fibroblast-coated collagen microtissues containing PHHs display high hepatic functions for 6+ weeks and are useful for assessing drug-mediated CYP induction and hepatotoxicity. Ultimately, microtissues may find utility for modeling liver diseases and as building blocks for cell-based therapies.

Original languageEnglish (US)
Pages (from-to)1-18
Number of pages18
JournalGene Expression The Journal of Liver Research
Volume20
Issue number1
DOIs
StatePublished - 2020

Bibliographical note

Publisher Copyright:
© 2020 Gene Expression The Journal of Liver Research. All rights reserved.

Keywords

  • 3T3-J2
  • Droplet microfluidics
  • Hepatic stellate cells (HSCs)
  • Spheroids

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