TY - JOUR
T1 - Regulation of hepatic stem/progenitor phenotype by microenvironment stiffness in hydrogel models of the human liver stem cell niche
AU - Lozoya, Oswaldo A.
AU - Wauthier, Eliane
AU - Turner, Rachael A.
AU - Barbier, Claire
AU - Prestwich, Glenn D.
AU - Guilak, Farshid
AU - Superfine, Richard
AU - Lubkin, Sharon R.
AU - Reid, Lola M.
N1 - Funding Information:
The Nucleic Acids Core Facility at UNC Chapel Hill provided oligonucleotide synthesis services for gene expression assays. The Histology Core Facility at UNC Chapel Hill provided cryosectioning services for the experiment. The Functional Genomics Core Facility at UNC Chapel Hill provided access to Real-Time PCR hardware. The Center for Gastrointestinal Biology and Diseases ImmunoTechnologies Core Facility at UNC Chapel Hill performed ELISA studies. All of the studies were financed by grants to LM Reid, including NIH grants ( AA014243 , IP30-DK065933 ), a Department of Energy Grant ( DE-FG02-02ER-63477 ) and sponsored research grants from Vesta Therapeutics (Bethesda, MD) and GigaCyte (Branford, CT). Support for SR Lubkin was provided in part by an NIH grant ( R01GM096195 ). In addition, funding for one semester of OA Lozoya’s salary and studies in R Superfine’s lab were provided by an NIH project grant ( 5-P41-EB002025 ). Studies in F Guilak’s lab were financed by an NIH grant ( AR50245 ). Studies for development of the thiol-modified HA derivatives were financed by an NIH grant ( DC04336 ) to GD Prestwich.
PY - 2011/10
Y1 - 2011/10
N2 - 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.
AB - 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.
KW - Differentiation
KW - Human hepatic stem cells
KW - Human hepatoblasts
KW - Hyaluronans
KW - Maturational lineages
KW - Microenvironment mechanical properties
UR - http://www.scopus.com/inward/record.url?scp=80051545803&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2011.06.042
DO - 10.1016/j.biomaterials.2011.06.042
M3 - Article
C2 - 21788068
AN - SCOPUS:80051545803
SN - 0142-9612
VL - 32
SP - 7389
EP - 7402
JO - Biomaterials
JF - Biomaterials
IS - 30
ER -