Functional differentiation of hepatocyte-like spheroid structures from putative liver progenitor cells in three-dimensional peptide scaffolds.
We investigated the ability of a new type of biological material, the self-assembling peptide scaffold, to foster tissue-like function by a putative adult rat hepatocyte progenitor cell line, Lig-8. In conventional adherent petri-dish cultures, Lig-8 cells divide exponentially, express markers for definitive endoderm HNF3 beta and hepatocyte lineage, including CK8 and alpha-fetoprotein, but lack expression of mature hepatocyte markers. However, in the three-dimensional peptide scaffold cultures, Lig-8 exhibits non-exponential cell kinetics, acquires a spheroidal morphology, and produces progeny cells with mature hepatocyte properties. The differentiated progeny cells display expression of transcription factor C/EBP alpha and several other indicators that suggest hepatocyte maturation, including binucleation, up-regulation of albumin, and expression of cytochrome P450s CYP1A1, CYP1A2, and CYP2E1. Moreover, all three cytochrome p450 enzyme activities are induced using 3-methylcholanthrene in these spheroids. These results suggest that a designed biological material may provide a conducive microenvironment in which putative adult progenitor cells differentiate into functional hepatocyte-like spheroid clusters. This bioengineered scaffold system provides a better physiological approach to "progenitor cell differentiation" for future biomedical and pharmaceutics applications.
Semino
Carlos E
CE
Center for Biomedical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA. semino@mit.edu
Merok
Joshua R
JR
Crane
Gracy G
GG
Panagiotakos
Georgia
G
Zhang
Shuguang
S
D000818:Animals; D001672:Biocompatible Materials; D002454:Cell Differentiation / Q000502:physiology*; D007700:Kinetics; D008099:Liver / Q000196:embryology*; D010455:Peptides; D051381:Rats; D013234:Stem Cells / Q000502:physiology*
D001672:Biocompatible Materials; D010455:Peptides