Functionally Enhanced Human Stem Cell Derived Hepatocytes in Galactosylated Cellulosic Sponges for Hepatotoxicity...

Area of Science:

• Biotechnology
• Stem Cell Biology
• Hepatology

Background:

• Pluripotent stem cell-derived hepatocyte-like cells (hPSC-HLCs) are a promising alternative to primary human hepatocytes (PHHs) for research and drug testing.
• Maintaining and improving the mature functions of hPSC-HLCs, especially for long-term applications, remains a significant challenge.
• 3D culture systems are being explored to support the formation and maintenance of functional hepatocyte spheroids for extended periods.

Purpose of the Study:

• To evaluate the efficacy of a 3D cellulosic scaffold system for culturing hPSC-HLCs.
• To assess the impact of the 3D scaffold on the maturation, functionality, and drug response of hPSC-HLCs.
• To determine the suitability of this 3D system for long-term maintenance and direct use in liver drug testing assays.

Main Methods:

• Human pluripotent stem cell-derived hepatocyte-like cells (hPSC-HLCs) were cultured within a macroporous 3D cellulosic scaffold.
• The scaffold's ability to support spheroid formation and maintenance over time was assessed.
• Hepatocyte functionality, including urea production, hepatic marker expression, and cytochrome P450 activity, was measured.
• The sensitivity of hPSC-HLCs in the scaffold to hepatotoxicants was compared to conventional 2D cultures and PHHs.

Main Results:

• The 3D cellulosic scaffold facilitated the formation and maintenance of optimal-sized hPSC-HLC spheroids for several weeks.
• Culturing in the 3D scaffold significantly enhanced hPSC-HLC functionality, evidenced by increased urea production and hepatic marker expression.
• hPSC-HLCs in the 3D scaffold exhibited a more mature phenotype with enhanced cytochrome P450 activity and induction.
• The system demonstrated higher sensitivity to hepatotoxicants and greater similarity to PHHs compared to 2D cultures.

Conclusions:

• 3D cellulosic scaffolds are effective for the long-term culture and maturation of hPSC-HLCs.
• The enhanced cellular function and mature phenotype in the 3D system make it highly suitable for drug testing applications.
• This technology provides a robust platform for maintaining mature hPSC-HLCs for direct use in hepatotoxicity assays, advancing drug safety evaluation.