title: Regulation of hepatic stem/progenitor phenotype by microenvironment stiffness in hydrogel models of the human liver stem cell niche.
abstract: Human livers have maturational lineages of cells within liver acini, beginning periportally in stem cell niches, the canals of Hering, and ending in polyploid hepatocytes pericentrally and cholangiocytes in bile ducts. Hepatic stem cells (hHpSCs) in vivo are partnered with mesenchymal precursors to endothelia (angioblasts) and stellate cells, and reside in regulated microenvironments, stem cell niches, containing hyaluronans (HA). The in vivo hHpSC niche is modeled in vitro by growing hHpSC in two-dimensional (2D) cultures on plastic. We investigated effects of 3D microenvironments, mimicking the liver's stem cell niche, on these hHpSCs by embedding them in HA-based hydrogels prepared with Kubota's Medium (KM), a serum-free medium tailored for endodermal stem/progenitors. The KM-HA hydrogels mimicked the niches, matched diffusivity of culture medium, exhibited shear thinning and perfect elasticity under mechanical loading, and had predictable stiffness depending on their chemistry. KM-HA hydrogels, which supported cell attachment, survival and expansion of hHpSC colonies, induced transition of hHpSC colonies towards stable heterogeneous populations of hepatic progenitors depending on KM-HA hydrogel stiffness, as shown by both their gene and protein expression profile. These acquired phenotypes did not show morphological evidence of fibrotic responses. In conclusion, this study shows that the mechanical properties of the microenvironment can regulate differentiation in endodermal stem cell populations.
authors.affiliation: Joint Department of Biomedical Engineering, North Carolina State University, Raleigh, NC and UNC School of Medicine, Chapel Hill, NC 27599, USA.
authors.forename: Oswaldo A
authors.initials: OA
authors.lastname: Lozoya
authors.forename: Eliane
authors.initials: E
authors.lastname: Wauthier
authors.forename: Rachael A
authors.initials: RA
authors.lastname: Turner
authors.forename: Claire
authors.initials: C
authors.lastname: Barbier
authors.forename: Glenn D
authors.initials: GD
authors.lastname: Prestwich
authors.forename: Farshid
authors.initials: F
authors.lastname: Guilak
authors.forename: Richard
authors.initials: R
authors.lastname: Superfine
authors.forename: Sharon R
authors.initials: SR
authors.lastname: Lubkin
authors.forename: Lola M
authors.initials: LM
authors.lastname: Reid
mesh_terms: D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D004058:Diffusion; D004548:Elasticity; D015870:Gene Expression; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000378:metabolism*; D020100:Hydrogels / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000166:cytology*; D010641:Phenotype; D055153:Stem Cell Niche; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D020100:Hydrogels; D006820:Hyaluronic Acid