title: The advantages of three-dimensional culture in a collagen hydrogel for stem cell differentiation.
abstract: We evaluated the advantages of three-dimensional (3D) culture in a collagen hydrogel for stem cell differentiation, including the morphology of differentiated cells, differentiation efficiency of stem cells from aged rat and cells after passaging and freeze/thawing. Rat mesenchymal stem cells (MSCs) from young and aged rats, and MSCs after passaging and freeze/thawing were induced to differentiate into osteoblasts in 3D and 2D cultures, and histological studies were performed. Differentiation efficiency was evaluated by markers of osteoblastic differentiation including Runx2 and osterix gene expressions, osteocalcin secretion and calcium deposition. MSCs were stained positive for alkaline phosphatase in 3D and 2D cultures. However, the morphology of differentiated cells in 3D culture, which was different from that in 2D culture, was similar to that of osteoblasts in vivo. Markers of osteoblastic differentiation in MSCs from aged rats in 3D culture were higher than those in MSCs from young rats in 2D culture. Markers of osteoblastic differentiation in MSCs after passaging and freeze/thawing in 3D culture were higher than those in nonpassaged MSCs in 2D culture. These results indicate that 3D culture in a collagen hydrogel has advantages for the differentiation of MSCs into osteoblasts with a similar phenotype to that of in vivo, when using even MSCs from aged donors or after passaging and freeze/thawing.
authors.affiliation: Department of Thoracic and Cardiovascular Surgery, Nara Medical University School of Medicine, Kashihara, Nara 634-8522, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Naito
authors.forename: Mamiko
authors.initials: M
authors.lastname: Yoshimura
authors.forename: Toshihide
authors.initials: T
authors.lastname: Mizuno
authors.forename: Shin
authors.initials: S
authors.lastname: Takasawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Tojo
authors.forename: Shigeki
authors.initials: S
authors.lastname: Taniguchi
mesh_terms: D000375:Aging / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects*; D003094:Collagen / Q000494:pharmacology*; D005615:Freezing; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051381:Rats
chemical_list: D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D003094:Collagen; D002118:Calcium
keywords: collagen hydrogel; differentiation efficiency; osteoblast; stem cell; three-dimensional culture