Automated Generation of hiPSC-Derived Hepatic Progeny by Cost-Efficient Compounds

Area of Science:

• Stem cell biology
• Hepatology
• Drug discovery and toxicology

Background:

• Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) show potential for liver disease modeling and drug screening.
• Current methods face challenges in cost-effectiveness and process automation.

Purpose of the Study:

• To develop a scalable, automated, and cost-efficient protocol for HLC differentiation from hPSCs.
• To identify chemical alternatives to expensive growth factors and extracellular matrices.

Main Methods:

• Utilized an automated liquid handling system for reproducible hPSC differentiation.
• Screened 92 chemicals to replace costly growth factors, achieving ~79% cost reduction.
• Evaluated recombinant extracellular matrices, specifically Laminin-521, as a Matrigel alternative.
• Optimized a small molecule combination for differentiation.

Main Results:

• Automated differentiation yielded highly reproducible HLCs.
• Small molecule optimization and Laminin-521 resulted in HLCs transcriptionally identical to growth factor-derived HLCs.
• Optimized HLCs exhibited comparable albumin/urea secretion, low AFP levels, similar CYP3A4 activity, and drug toxicity profiles.
• The protocol was validated across four different hPSC lines.

Conclusions:

• Developed a scalable, xeno-free, and cost-efficient method for producing hPSC-derived HLCs.
• The optimized protocol is suitable for high-throughput disease modeling, drug screening, and potentially regenerative therapies.