Hepatic differentiation of human iPSCs in different 3D models: a comparative study

Florian Meier, Nora Freyer, Joanna Brzeszczynska, Fanny Knöspel, Lyle Armstrong, Majlinda Lako, Selina Greuel, Georg Damm, Eva Ludwig-Schwellinger, Ulrich Deschl, James A. Ross, Mario Beilmann, Katrin Zeilinger

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)

Abstract

Human induced pluripotent stem cells (hiPSCs) are a promising source from which to derive distinct somatic cell types for in vitro or clinical use. Existent protocols for hepatic differentiation of hiPSCs are primarily based on 2D cultivation of the cells. In the present study, the authors investigated the generation of hiPSC-derived hepatocyte-like cells using two different 3D culture systems: A 3D scaffold-free microspheroid culture system and a 3D hollow-fiber perfusion bioreactor. The differentiation outcome in these 3D systems was compared with that in conventional 2D cultures, using primary human hepatocytes as a control. The evaluation was made based on specific mRNA expression, protein secretion, antigen expression and metabolic activity. The expression of α-fetoprotein was lower, while cytochrome P450 1A2 or 3A4 activities were higher in the 3D culture systems as compared with the 2D differentiation system. Cells differentiated in the 3D bioreactor showed an increased expression of albumin and hepatocyte nuclear factor 4α, as well as secretion of α-1-antitrypsin as compared with the 2D differentiation system, suggesting a higher degree of maturation. In contrast, the 3D scaffold-free microspheroid culture provides an easy and robust method to generate spheroids of a defined size for screening applications, while the bioreactor culture model provides an instrument for complex investigations under physiological-like conditions. In conclusion, the present study introduces two 3D culture systems for stem cell derived hepatic differentiation each demonstrating advantages for individual applications as well as benefits in comparison with 2D cultures.
Original languageEnglish
Pages (from-to)1759-1771
Number of pages13
JournalInternational Journal of Molecular Medicine
Volume40
Issue number6
Early online date16 Oct 2017
DOIs
Publication statusE-pub ahead of print - 16 Oct 2017
Externally publishedYes

Keywords

  • human induced pluripotent stem cells
  • hepatocyte-like cells
  • 3D culture
  • primary human hepatocytes
  • hepatic differentiation

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