title: Chondrogenic differentiation of human mesenchymal stem cells in collagen type I hydrogels.
abstract: The chondrogenic differentiation of bone marrow-derived human mesenchymal stem cells (MSCs) in a collagen type I hydrogel, which is in clinical use for matrix-based autologous chondrocyte transplantation (ACT), was investigated. Collagen hydrogels with 2.5 x 10(5) MSCs/mL were fabricated and cultured for 3 weeks in a serum-free, defined, chondrogenic differentiation medium containing 10 ng/mL TGF-beta1 or 100 ng/mL BMP-2. Histochemistry revealed morphologically distinct, chondrocyte-like cells, surrounded by a sulfated proteoglycan-rich extracellular matrix in the TGF-beta1 and BMP-2 treated group, with more elongated cells seen in the BMP-2 treated group. Immunohistochemistry detected collagen type II (Col II) in the TGF-beta1 and BMP-2 treated group. Collagen type X (Col X) staining was positive in the TGF-beta1 but only very weak in the BMP-2 treated group. RT-PCR analyses revealed a specific chondrogenic differentiation with the expression of the cartilage specific marker genes Col II, Col X, and aggrecan (AGN) in the TGF-beta1 and the BMP-2 treated group, with earlier expression of these marker genes in the TGF-beta1 treated group. Interestingly, MSC-gels cultured in DMEM with 10% FBS (control) indicated few isolated chondrocyte-like cells but no expression of Col II or Col X could be detected. The results show, that MSCs cultured in a collagen type I hydrogel are able to undergo a distinct chondrogenic differentiation pathway, similar to that described for MSCs cultured in high-density pellet cultures. These findings are valuable in terms of ex vivo predifferentiation or in situ differentiation of MSCs in collagen hydrogels for articular cartilage repair.
authors.affiliation: University of Würzburg, Orthopedic Center for Musculoskeletal Research, König-Ludwig-Haus, Brettreichstr. 11, 97070 Würzburg, Germany. u-noeth.klh@mail.uni-wuerzburg.de
authors.forename: Ulrich
authors.initials: U
authors.lastname: Nöth
authors.forename: Lars
authors.initials: L
authors.lastname: Rackwitz
authors.forename: Andrea
authors.initials: A
authors.lastname: Heymer
authors.forename: Meike
authors.initials: M
authors.lastname: Weber
authors.forename: Bernd
authors.initials: B
authors.lastname: Baumann
authors.forename: Andre
authors.initials: A
authors.lastname: Steinert
authors.forename: Norbert
authors.initials: N
authors.lastname: Schütze
authors.forename: Franz
authors.initials: F
authors.lastname: Jakob
authors.forename: Jochen
authors.initials: J
authors.lastname: Eulert
mesh_terms: D000943:Antigens, Differentiation / Q000096:biosynthesis; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D024042:Collagen Type I / Q000737:chemistry; D024043:Collagen Type II / Q000096:biosynthesis; D024121:Collagen Type X / Q000096:biosynthesis; D005260:Female; D006801:Humans; D020100:Hydrogels; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D014182:Transplantation, Autologous
chemical_list: D000943:Antigens, Differentiation; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; D024043:Collagen Type II; D024121:Collagen Type X; D020100:Hydrogels; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1