Substrate-dependent Wnt signaling in MSC differentiation within biomaterial-derived 3D spheroids
Journal
Biomaterials
Journal Volume
34
Journal Issue
20
Pages
4725-4738
Date Issued
2013
Author(s)
Abstract
A unique biomaterial-based system was developed to generate dynamic three-dimensional (3D) multicellular spheroids of mesenchymal stem cells (MSCs). MSCs were cultured on transparent membranes made of chitosan or those further grafted with hyaluronan (HA) in different densities. MSCs vigorously migrated and were self-assembled into highly mobile 3D spheroids with substrate-dependent upregulation of adhesion molecule N-cadherin. MSC spheroids showed increased expression of Wnt genes/proteins and substrate-dependent cell fate. The correlation of differentiation capacities with Wnt signaling and crosstalk with other pathways such as ERK1/2 or Smad2/3 were observed for MSC spheroids but not for the conventional 2D cultured cells. Wnt3a-mediated canonical Wnt signaling was more active for MSC spheroids derived on chitosan, which were prone to osteogenesis. Wnt5a-mediated non-canonical Wnt signaling was more active for MSC spheroids derived on HA-grafted chitosan, which were prone to chondrogenesis. In particular, the relative importance of Wnt5a-mediated non-canonical vs. Wnt3a-mediated canonical Wnt signals in determining the cell fate was controlled by the grafting density of HA on chitosan. Treatment with the inhibitor of canonical Wnt-associated signaling molecules suppressed the osteogenesis of MSC spheroids on chitosan. This study demonstrates that Wnt signaling of MSCs is distinct in 3D environment and is substrate-dependent. The convenient 3D platform may be used to examine the role of Wnt signaling in controlling MSC fate under different extracellular environments, and potentially applied to study stem cell behavior in regenerative medicine, normal development, and cancer. ? 2013 Elsevier Ltd.
Subjects
Chitosan; Hyaluronan; Mesenchymal stem cells (MSCs); Spheroids; Wnt signaling
SDGs
Other Subjects
Canonical Wnt signaling; Extracellular environments; Hyaluronan; Mesenchymal stem cell; Multicellular spheroid; Regenerative medicine; Spheroids; Wnt signaling; Biological materials; Biomaterials; Chitosan; Flowcharting; Grafting (chemical); Hyaluronic acid; Molecules; Signaling; Stem cells; Substrates; Three dimensional computer graphics; Cell culture; biomaterial; chitosan; hyaluronic acid; mitogen activated protein kinase 1; mitogen activated protein kinase 3; nerve cell adhesion molecule; Smad2 protein; Smad3 protein; Wnt protein; Wnt5a protein; animal cell; animal experiment; animal tissue; article; bone development; cell adhesion; cell culture; cell density; cell differentiation; cell fate; chondrogenesis; controlled study; enzyme substrate; gene expression; mesenchymal stem cell; nonhuman; priority journal; rat; signal transduction; spheroid cell; upregulation; Animals; Biocompatible Materials; Cadherins; Cell Differentiation; Cell Movement; Cell Shape; Chondrogenesis; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Glycosaminoglycans; Mesenchymal Stromal Cells; Models, Biological; Osteogenesis; Rats; Rats, Sprague-Dawley; Signal Transduction; Smad Proteins; Spheroids, Cellular; Wnt Proteins
Type
journal article