Chondrogenic differentiation of human mesenchymal stem cells in collagen type I hydrogels

Nöth, Ulrich; Rackwitz, Lars; Heymer, A; Weber, M.; Baumann, Bernd; Steinert, Andre F.; Schütze, Norbert; Jakob, Franz; Eulert, J.

doi:10.1002/jbm.a.31254

Cited by 171 publications

(123 citation statements)

References 45 publications

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“…For a long time, BMP-2 has been used as a strong stimulator for osteogenic differentiation of stem cells [35]. In other studies, however, BMP-2 has been applied to promote the chondrogenic differentiation of MSCs as well [24,26]. Meanwhile, it is also believed that BMP-2-induced chondrogenic differentiation of MSCs will eventually lead to hypertrophy and endochondral ossification [36,37].…”

Section: Discussionmentioning

confidence: 99%

“…In line with increased EdU incorporation in co-culture pellets, 1 cluster contained intracellular cell-cycle regulators, such as CCND1, -2, and -3. The fourth cluster contained two secreted growth factors FGF-1 and BMP-2, both of which are established modulators of chondrocyte proliferation and/or matrix production [24][25][26].…”

Section: Microarray Study Identifies a Group Of Genes Regulated By Thmentioning

confidence: 99%

See 1 more Smart Citation

Fibroblast Growth Factor-1 Is a Mesenchymal Stromal Cell-Secreted Factor Stimulating Proliferation of Osteoarthritic Chondrocytes in Co-Culture

Leijten

Blitterswijk

et al. 2013

Stem Cells and Development

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Previously, we showed that mesenchymal stromal cells (MSCs) in co-culture with primary chondrocytes secrete soluble factors that increase chondrocyte proliferation. The objective of this study is to identify these factors. Human primary chondrocytes (hPCs) isolated from late-stage osteoarthritis patients were co-cultured with human bone marrow-derived MSCs (hMSCs) in pellets. Genome-wide mRNA expression analysis and quantitative polymerase chain reactions (qPCR) were used to identify soluble factors that were specifically induced in co-cultures. Immunofluorescent staining combined with cell tracking and enzyme-linked immunosorbent assay (ELISA) were performed to validate up-regulation at the protein level and to identify the cellular origin of the increased proteins. Chemical blockers and neutralizing antibodies were used to elucidate the role of the identified candidate genes in co-cultures. A number of candidate factors were differentially regulated in co-cultures at the mRNA level. Of these, fibroblast growth factor-1 (FGF-1) mRNA and protein expression were markedly increased in co-cultures predominantly due to up-regulated expression in MSCs. Blocking of FGF signaling in co-culture pellets by specific FGF receptor inhibitors or FGF-1 neutralizing antibodies completely blocked hPCs proliferation. We demonstrate that MSCs increase FGF-1 secretion on co-culture with hPCs, which, in turn, is responsible for increased hPCs proliferation in pellet co-cultures. C artilage repair is among the key targets of regenerative medicine. Several stem cell-based therapies have been proposed to treat cartilage defects, rheumatic arthritis, osteoarthiris (OA), and other joint diseases. These therapeutic strategies include implantation or injection of mesenchymal stromal cells (MSCs) in the diseased joint or bone marrow stimulation techniques such as micro fracture. These procedures rely on cellular interactions between MSCs and native cells in the joint [1][2][3][4]. Recent studies showed that intra-articularly injected MSCs can reduce incidence and severity of arthritis in animal model by inducing immune tolerance [5]. Further investigations showed that MSCs are capable of inducing hyporesponsiveness of T-lymphocytes and of expressing antiinflammatory mediators in macrophages [6,7]. Besides immuno-suppressive effects, intra-articularly injected MSCs were also reported to inhibit thickening of the synovium and to protect against cartilage destruction in a mouse OA model [8]. However, with the cellular interactions between MSCs and native chondrocytes receiving less attention [9], the molecular mechanism of interactions between MSCs and native chondrocytes is poorly understood [10]. In order to maximize the benefits of stem cell therapy, these cellular interactions need to be clarified.Previous reports showed that cartilage matrix formation was increased in co-cultures of human primary chondrocytes (hPCs) with MSCs compared with monocultures [11,12]. Further investigations revealed that the inductive effects of MSCs on cartila...

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Section: Discussionmentioning

confidence: 99%

Section: Microarray Study Identifies a Group Of Genes Regulated By Thmentioning

confidence: 99%

Fibroblast Growth Factor-1 Is a Mesenchymal Stromal Cell-Secreted Factor Stimulating Proliferation of Osteoarthritic Chondrocytes in Co-Culture

Leijten

Blitterswijk

et al. 2013

Stem Cells and Development

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“…However, Nöth et al reported a contraction of up to 75% in the collagen gels, a process that could be results can be confirmed in in vivo studies, the application of MSCs will be facilitated. Our study does not prodisadvantageous for implantation purposes (33). We chose a low cell concentration for differentiation vide new insights regarding this aspect, but the gelatin hydrogels were found to be a suitable material for either due to the limited nutrient supply expected within the intervertebral disc tissue.…”

Section: Establishing Hypoxic Conditions and Experimental Setup Measumentioning

confidence: 99%

“…The gelatin hydrogels presented here as a model for the preparation of tion within a gelatin matrix. (33,54). However, Nöth et al reported a contraction of up to 75% in the collagen gels, a process that could be results can be confirmed in in vivo studies, the application of MSCs will be facilitated.…”

Section: Establishing Hypoxic Conditions and Experimental Setup Measumentioning

confidence: 99%

Hypoxic Conditions during Expansion Culture Prime Human Mesenchymal Stromal Precursor Cells for Chondrogenic Differentiation in Three-Dimensional Cultures

et al. 2011

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Cell-based approaches using mesenchymal stromal precursor cells (MSCs) for the regeneration of intervertebral discs are attracting increased interest, even though the intervertebral disc is a very demanding environment. Implanted cells eventually face acidic pH, hypoxia, and a lack of nutrients. While the regenerative potential of MSCs for skeletal tissues has been well described, it is still questionable whether human MSCs can be prepared for prolonged survival and proper functioning and whether they can differentiate under the adverse conditions encountered in the disc. Here we examined the influence of hypoxia during expansion and differentiation on the chondrogenesis of MSCs. Chondrogenic differentiation was performed in in situ solidifying gelatin hydrogels, which represent a suitable matrix for delivering and anchoring cells within the disc tissue. To consider limitations in nutrition in the intervertebral disc, differentiation was performed at low cell concentrations in the gelatin hydrogels. Standard high-density micromass cultures served as reference controls. To determine the quality of chondrogenesis we analyzed typical marker molecules such as collagen types I, II, X, Sox-9, MIA, and aggrecan mRNA using RT-qPCR and determined protein deposition by histological stainings and biochemical methods. We could demonstrate that in gelatin-based hydrogels chondrogenic differentiation of human MSCs is possible at low cell concentrations. The quality of chondrogenic differentiation could be improved by hypoxia. Best results were obtained when the entire in vitro process, including MSC expansion and subsequent differentiation, was done under hypoxic conditions. MSCs that were expanded under reduced oxygen tension were primed for a chondrogenic differentiation.

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“…Attempts to mimic this environment by using hydrogel materials has shown promise in restoring functionally competent cartilage because of the inherent cross-linked hydrogel polymer networks which facilitate efficient transport of nutrients and actively form physical environments similar to that of articular cartilage [17]. Natural hydrogel polymers include chitosan, collagen, hyaluronic acid, gelatin and alginate, whereas synthetic polymers encompass poly(glycolic acid) (PGA), poly(lactic acid) (PLA), poly(ethylene oxide) (PEO), poly(vinyl alcohol) (PVA), poly(acrylic acid) (PAA) and their copolymers [18][19][20][21][22][23][24]. Typically, such hydrogel materials can be functionalized or assembled with specific functional groups to impart enhanced biocompatible properties.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Engineered Peptide-Glycosaminoglycan Microfibrous Hybrid Scaffolds for Potential Applications in Cartilage Tissue Regeneration

Romanelli

Knoll

Santora

et al. 2015

Fibers

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Advances in tissue engineering have enabled the ability to design and fabricate biomaterials at the nanoscale that can actively mimic the natural cellular environment of host tissue. Of all tissues, cartilage remains difficult to regenerate due to its avascular nature. Herein we have developed two new hybrid polypeptide-glycosaminoglycan microfibrous scaffold constructs and compared their abilities to stimulate cell adhesion, proliferation, sulfated proteoglycan synthesis and soluble collagen synthesis when seeded with chondrocytes. Both constructs were designed utilizing self-assembled Fmoc-protected valyl cetylamide nanofibrous templates. The peptide components of the constructs were varied. For Construct I a short segment of dentin sialophosphoprotein followed by Type I collagen were attached to the templates using the layer-by-layer approach. For Construct II, a short peptide segment derived from the integrin subunit of Type II collagen binding protein expressed by chondrocytes was attached to the templates followed by Type II collagen. To both constructs, we then attached the natural polymer N-acetyl glucosamine, chitosan. Subsequently, the glycosaminoglycan chondroitin sulfate was then attached as the final layer. The scaffolds were characterized by Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), atomic force microscopy and scanning electron microscopy. In vitro culture studies were carried out in the presence of chondrocyte cells for OPEN ACCESSFibers 2015, 3 266 both scaffolds and growth morphology was determined through optical microscopy and scanning electron microscopy taken at different magnifications at various days of culture. Cell proliferation studies indicated that while both constructs were biocompatible and supported the growth and adhesion of chondrocytes, Construct II stimulated cell adhesion at higher rates and resulted in the formation of three dimensional cell-scaffold matrices within 24 h. Proteoglycan synthesis, a hallmark of chondrocyte cell differentiation, was also higher for Construct II compared to Construct I. Soluble collagen synthesis was also found to be higher for Construct II. The results of the above studies suggest that scaffolds designed from Construct II be superior for potential applications in cartilage tissue regeneration. The peptide components of the constructs play an important role not only in the mechanical properties in developing the scaffolds but also control cell adhesion, collagen synthesis and proteoglycan synthesis capabilities.

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Chondrogenic differentiation of human mesenchymal stem cells in collagen type I hydrogels

Cited by 171 publications

References 45 publications

Fibroblast Growth Factor-1 Is a Mesenchymal Stromal Cell-Secreted Factor Stimulating Proliferation of Osteoarthritic Chondrocytes in Co-Culture

Fibroblast Growth Factor-1 Is a Mesenchymal Stromal Cell-Secreted Factor Stimulating Proliferation of Osteoarthritic Chondrocytes in Co-Culture

Hypoxic Conditions during Expansion Culture Prime Human Mesenchymal Stromal Precursor Cells for Chondrogenic Differentiation in Three-Dimensional Cultures

Comparison of Engineered Peptide-Glycosaminoglycan Microfibrous Hybrid Scaffolds for Potential Applications in Cartilage Tissue Regeneration

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