# High-throughput exploration of chemomechanical crosstalk in the maturation of iPSC-derived human hepatocytes

> **NIH NIH R21** · UNIVERSITY OF ILLINOIS AT CHICAGO · 2020 · $184,392

## Abstract

PROJECT SUMMARY / ABSTRACT
Differences in drug metabolism pathways across species necessitate the use of in vitro human liver models for
screening the metabolism and toxicity of drugs/industrial chemicals, and for discovering novel therapeutics
against liver diseases. While primary human hepatocytes (PHHs) are the ‘gold standard’ for fabricating human
liver models, a severe shortage of donor livers limits their utility for high-throughput screening and prevents a
comprehensive assessment of genetic determinants of diseases. In contrast, induced pluripotent stem cell-
derived human hepatocyte-like cells (iPSC-HHs) created from genetically diverse donor panels can address
the limitations of PHHs; however, the protocols to generate iPSC-HHs cannot fully mature these cells towards
an adult PHH phenotype and thus such protocols require substantial refinement. An enhanced understanding
of how liver-inspired microenvironmental cues synergistically affect iPSC-HH maturation and maintenance of
phenotype is urgently needed to utilize iPSC-HHs more effectively for the above applications. The adult liver
contains an extracellular matrix (ECM) that a) presents biochemical and biomechanical signals, and b) can
further modulate interactions of liver cells with growth factors and cytokines. Previous investigations have
demonstrated that the composition of ECM, substrate stiffness, and growth factors can independently influence
the in vitro functions of primary hepatocytes and embryonic stem cell-derived hepatocyte-like cells. However,
the crosstalk between these cues as occurs in vivo is unclear, and the combinatorial effects of ECM and
growth factor signals on the functional maturation of iPSC-HHs have yet to be identified. In addition, the liver
has several non-parenchymal cell (NPC) types that can influence hepatocellular functions, including liver
sinusoidal endothelial cells (LSECs) that provide critical regulatory signals to hepatocytes during liver
development, physiology, and regeneration; our recent work also supports the notion that LSECs regulate
iPSC-HH functions to a greater degree than other liver NPC types. Thus, in this proposal we will examine the
central hypothesis that ECM composition/stiffness and LSEC intercellular interactions collectively act to
significantly modulate iPSC-HH functional maturity. Part of the challenge in testing such a hypothesis is that
evaluating a large number of combinations of microenvironmental cues in a bulk culture format is too costly as
well as labor and time intensive. Therefore, we will adapt a cellular microarray platform that enables defined
ECM microenvironments, combinatorial culture with soluble factors, and quantitative assessment of cell
phenotype using high-content imaging readouts. In Aim 1, we will systematically investigate the cooperative
effects of ECM composition, substrate stiffness, and growth factor signaling on iPSC-HH phenotype. In Aim 2,
we will examine reciprocal interactions between iPSC-H...

## Key facts

- **NIH application ID:** 10022330
- **Project number:** 5R21ES028580-02
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT CHICAGO
- **Principal Investigator:** Salman R Khetani
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $184,392
- **Award type:** 5
- **Project period:** 2019-09-23 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10022330

## Citation

> US National Institutes of Health, RePORTER application 10022330, High-throughput exploration of chemomechanical crosstalk in the maturation of iPSC-derived human hepatocytes (5R21ES028580-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10022330. Licensed CC0.

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