title: The effects of covalently immobilized hyaluronic acid substrates on the adhesion, expansion, and differentiation of embryonic stem cells for in vitro tissue engineering.
abstract: We investigated the in vitro effects of the molecular weight (MW) of hyaluronic acid (HA) on the maintenance of the pluripotency and proliferation of murine embryonic stem (ES) cells. High (1000 kDa) or low (4-8 kDa) MW HA was derivatized using an ultraviolet-reactive compound, 4-azidoaniline, and the derivative was immobilized onto cell culture cover slips. Murine ES cells were cultured on these HA surfaces for 5 days. High-MW HA interacted with murine ES cells via CD44, whereas low-MW HA interacted with these cells mostly via CD168. ES cells grown on both high- and low-MW HA appeared undifferentiated after 3 days. However, more cells adhered, proliferated, and exhibited greater amounts of phospho-p42/44 mitogen-activated-protein-kinase on low- compared with high-MW HA. Expression of Oct-3/4 and phosphorylation of STAT3 were enhanced by ES cells on low-MW HA, not on high-MW HA. After release from HA, cells cultured on low-MW HA in the presence of differentiating medium showed enhanced expression of α-SMA or CD31 compared with cells cultured on high-MW HA. It was concluded that low-MW HA substrates were effective in maintaining murine ES cells in a viable and undifferentiated state, which favors their use in the propagation of ES cells for tissue engineering.
authors.affiliation: Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, 2-1-Hirosawa, Wako-shi, Saitama 351-0198, Japan. b-joddar@riken.jp
authors.forename: Binata
authors.initials: B
authors.lastname: Joddar
authors.forename: Takashi
authors.initials: T
authors.lastname: Kitajima
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ito
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D017931:DNA Primers; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000201:enzymology; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000378:metabolism; D006820:Hyaluronic Acid / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D008970:Molecular Weight; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D050796:STAT3 Transcription Factor / Q000378:metabolism; D023822:Tissue Engineering
chemical_list: C476399:Cd44 protein, mouse; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D018960:Hyaluronan Receptors; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; C098946:hyaluronan-mediated motility receptor; D006820:Hyaluronic Acid; D000469:Alkaline Phosphatase