1 research outputs found
Substrate Stiffness Modulates the Maturation of Human Pluripotent Stem-Cell-Derived Hepatocytes
Obtaining functional hepatocytes
from human pluripotent stem cells (hPSCs) holds great potential for
applications in drug safety testing, as well in the field of regenerative
medicine. However, developing functionally mature hPSC-derived hepatocytes
(hPSC-Heps) remains a challenge. We hypothesized that the cellular
microenvironment plays a vital role in the maturation of immature
hepatocytes. In this study, we examined the role of mechanical stiffness,
a key component of the cellular microenvironment, in the maturation
of hPSC-Heps. We cultured hPSC-Heps on collagen-coated polyacrylamide
hydrogels with varying elastic moduli. On softer substrates the hPSC-Heps
formed compact colonies while on stiffer substrates they formed a
diffuse monolayer. We observed an inverse correlation between albumin
production and substrate stiffness. The expression of key cytochrome
enzymes, which are expressed at higher levels in the adult liver compared
to the fetal liver, also correlated inversely with substrate stiffness,
whereas fetal markers such as Cyp3A7 and AFP showed no correlation
with stiffness. Culture of hPSC-Heps on soft substrates for 12 days
led to 10–30 fold increases in the expression of drug-metabolizing
enzymes. These results demonstrate that substrate stiffness similar
to that of the liver enables aspects of the maturation of hPSC-Heps