A chitosan-hyaluronan-based hydrogel-hydrocolloid supports in vitro culture and differentiation of human mesenchymal stem/stromal cells.
Three-dimensional (3D) cell culture platforms are increasingly utilized due to their ability to more closely mimic the in vivo microenvironment compared to traditional two-dimensional methods. Limitations of currently available 3D materials include lack of cell attachment, long polymerization times, and inclusion of undefined xenobiotics, and cytotoxic cross-linkers. Evaluated here is a unique hydrogel comprised of polyelectrolytic complex (PEC) fibers formed by hyaluronic acid and chitosan (CT). When hydrated with fetal bovine serum containing human mesenchymal stem/stromal cells (hMSCs), a hydrogel with an elastic modulus of 264±38 Pa formed in seconds with cells distributed throughout the matrix. Scanning electron microscopy showed a lattice-like meshwork of PEC fibers forming irregular compartments. hMSCs showed 48% viability during the first 24 h, with cell populations thereafter reaching a steady state for 14 days. hMSCs in the matrix were induced to differentiate to chondrogenic, osteogenic, and adipogenic phenotypes. Emergent features, at days 56 and 70, consisted of chondrogenesis on the surface of hydrogels induced to osteogenic and adipogenic phenotypes. Results indicate that this matrix may be useful for tissue engineering and disease modeling applications.
1Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota.|2Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota.|3Bioactive Regenerative Therapeutics, Inc., Duluth, Minnesota.
Beth A
BA
Lindborg
3Bioactive Regenerative Therapeutics, Inc., Duluth, Minnesota.
John H
JH
Brekke
4Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota.
Carolyn M
CM
Scott
3Bioactive Regenerative Therapeutics, Inc., Duluth, Minnesota.
Yi Wen
YW
Chai
1Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota.|2Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota.
Connor
C
Ulrich
5Department of Surgery, Section of Neurosurgery, St. John Providence Health System, Southfield, Michigan.
Lee
L
Sandquist
6Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota.
Efrosini
E
Kokkoli
1Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota.|2Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota.
Timothy D
TD
O'Brien
D017667:Adipocytes / Q000166:cytology; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002454:Cell Differentiation; D060833:Cellular Microenvironment; D048271:Chitosan; D019902:Chondrocytes / Q000166:cytology; D003102:Colloids; D055119:Elastic Modulus; D006801:Humans; D006820:Hyaluronic Acid; D020100:Hydrogels; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008855:Microscopy, Electron, Scanning; D010011:Osteocytes / Q000166:cytology; D010641:Phenotype; D012212:Rheology; D023822:Tissue Engineering / Q000295:instrumentation*; D014783:Viscosity
D003102:Colloids; D020100:Hydrogels; D006820:Hyaluronic Acid; D048271:Chitosan