THE CHALLENGE
The field of regenerative medicine and drug development urgently requires reliable in vitro liver models that accurately mimic primary human hepatocyte functions. Primary hepatocytes, though ideal for studying liver physiology, suffer from limited availability and high inter-donor variability. Induced pluripotent stem cells (iPSCs) hold promise as alternative sources but typically generate hepatocyte-like cells that remain functionally immature, retaining fetal characteristics that compromise their effectiveness in disease modeling and pharmaceutical testing. Current maturation techniques are fraught with challenges as they largely depend on high concentrations of exogenous chemicals or non-physiological culture conditions. These methods can lead to receptor oversaturation and activation of off-target signaling pathways, ultimately resulting in unstable cellular phenotypes. Additionally, conventional two-dimensional cultures or rudimentary scaffold systems fail to replicate the spatial and biochemical complexity of the native liver environment. This gap in current approaches underscores the critical need for innovative strategies that promote complete and stable hepatocyte maturation without relying on disruptive external modulators.
OUR SOLUTION
This invention presents a novel 3D organoid system for maturing induced pluripotent stem cell–derived hepatocyte-like cells by co-culturing them with liver sinusoidal endothelial cells and Kupffer cells in a defined extracellular matrix composed of collagen Type I and fibronectin at specific ratios. Unlike traditional methods that rely on high concentrations of exogenous chemicals, this approach leverages natural intercellular signaling—mediated by factors such as hepatocyte growth factor, prostaglandin E2, and oncostatin M—to transition cells from a fetal to a mature adult phenotype, as evidenced by decreased AFP and HNF-4a levels and increased albumin and cytochrome P450 activities. By mimicking the in vivo hepatic environment more closely, the system offers a repeatable and physiologically relevant method to generate functional hepatocytes identical to primary human hepatocytes, thereby overcoming the limitations of existing chemical-based maturation protocols.
Advantages:
- 3D organoid mimics liver microenvironment
- Functional maturation of hepatocyte-like cells
- Minimal use of exogenous chemicals
- Applications in drug testing and personalized medicine
Potential Application:
- Personalized medicine
- Drug discovery
- Toxicology screening
- Pharmaceutical testing
