Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: influence of collagen type II extracellular matrix on MSC chondrogenesis.
Bone marrow mesenchymal stem cells (MSCs) are candidate cells for cartilage tissue engineering. This is due to their ability to undergo chondrogenic differentiation after extensive expansion in vitro and stimulation with various biomaterials in three-dimensional (3-D) systems. Collagen type II is one of the major components of the hyaline cartilage and plays a key role in maintaining chondrocyte function. This study aimed at analyzing the MSC chondrogenic response during culture in different types of extracellular matrix (ECM) with a focus on the influence of collagen type II on MSC chondrogenesis. Bovine MSCs were cultured in monolayer as well as in alginate and collagen type I and II hydrogels, in both serum free medium and medium supplemented with transforming growth factor (TGF) beta1. Chondrogenic differentiation was detected after 3 days of culture in 3-D hydrogels, by examining the presence of glycosaminoglycan and newly synthesized collagen type II in the ECM. Differentiation was most prominent in cells cultured in collagen type II hydrogel, and it increased in a time-dependent manner. The expression levels of the of chondrocyte specific genes: sox9, collagen type II, aggrecan, and COMP were measured by quantitative "Real Time" RT-PCR, and genes distribution in the hydrogel beads were localized by in situ hybridization. All genes were upregulated by the presence of collagen, particularly type II, in the ECM. Additionally, the chondrogenic influence of TGF beta1 on MSCs cultured in collagen-incorporated ECM was analyzed. TGF beta1 and dexamethasone treatment in the presence of collagen type II provided more favorable conditions for expression of the chondrogenic phenotype. In this study, we demonstrated that collagen type II alone has the potential to induce and maintain MSC chondrogenesis, and prior interaction with TGF beta1 to enhance the differentiation.
Laboratory of Veterinary Surgery, Department of Clinical Science, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan. Darko.Bosnakovski@UTSouthwestern.edu
Darko
D
Bosnakovski
Morimichi
M
Mizuno
Gonhyung
G
Kim
Satoshi
S
Takagi
Masahiro
M
Okumura
Toru
T
Fujinaga
D053674:Aggrecans; D000464:Alginates / Q000494:pharmacology; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020219:Chondrogenesis / Q000187:drug effects / Q000502:physiology; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics; D024042:Collagen Type I / Q000235:genetics / Q000494:pharmacology; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D024121:Collagen Type X / Q000235:genetics; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005109:Extracellular Matrix / Q000502:physiology; D016326:Extracellular Matrix Proteins / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D020723:Glucuronic Acid / Q000494:pharmacology; D006023:Glycoproteins / Q000235:genetics; D006603:Hexuronic Acids / Q000494:pharmacology; D006609:High Mobility Group Proteins / Q000235:genetics; D020100:Hydrogels / Q000494:pharmacology*; D017403:In Situ Hybridization; D037181:Lectins, C-Type / Q000235:genetics; D064235:Matrilin Proteins; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D011506:Proteins / Q000494:pharmacology; D055758:SOX9 Transcription Factor; D023822:Tissue Engineering / Q000379:methods; D014157:Transcription Factors / Q000235:genetics
D053674:Aggrecans; D000464:Alginates; D011508:Chondroitin Sulfate Proteoglycans; D024042:Collagen Type I; D024043:Collagen Type II; D024121:Collagen Type X; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D006603:Hexuronic Acids; D006609:High Mobility Group Proteins; D020100:Hydrogels; D037181:Lectins, C-Type; D064235:Matrilin Proteins; D011506:Proteins; D055758:SOX9 Transcription Factor; D014157:Transcription Factors; D020723:Glucuronic Acid