Effects of Three-Dimensional Culture and Growth Factors on the Chondrogenic Differentiation of Murine Embryonic Stem Cells

Hwang, Nathaniel S.; Kim, Myoung Sook; Sampattavanich, Somponnat; Baek, Jin Hyen; Zhang, Zijun; Elisseeff, Jennifer H.

doi:10.1634/stemcells.2005-0024

Cited by 231 publications

(179 citation statements)

References 37 publications

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“…HUVECs (Cambrex, Walkersville, MD, passage 11-12) were cultured in an endothelial cell growth medium (Cambrex). NIH/3T3 mouse fibroblasts (passages [24][25][26] were cultured in DMEM (Mediatech, VA) supplemented with 5% cosmic calf serum (HyClone) and 3.7 g/l NaHCO 3 (ACROS Organics, NJ). HL-1 cells were cultured in Complete Claycomb Medium (SAFC Biosciences, KS).…”

Section: Biological Characterization Of Hydrogel Arraysmentioning

confidence: 99%

“…PEG chains derivatized on each end with vinyl-containing functional groups have been a particularly prevalent base polymer, as they can be readily processed into a cell-laden hydrogel network via photo-crosslinking [7,19] and can be derivatized via simple chemistries to generate networks that contain ECM-derived signals [20,21]. These properties have led to widespread use of PEG hydrogels as matrices to support tissue formation by various cell types, including osteoblasts [22], chondrocytes [23,24], neural cells [6], and multiple stem cell types [7,17,25,26]. For example, in a series of studies Anseth and coworkers have used PEG-based hydrogels to demonstrate that 3-D cartilage formation is influenced by ECM chemistry [27], mechanics [23], and degradation [28,29].…”

Section: Introductionmentioning

confidence: 99%

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An adaptable hydrogel array format for 3-dimensional cell culture and analysis

et al. 2008

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Hydrogels have been commonly used as model systems for 3-dimensional (3-D) cell biology, as they have material properties that resemble natural extracellular matrices (ECMs), and their cellinteractive properties can be readily adapted in order to address a particular hypothesis. Natural and synthetic hydrogels have been used to gain fundamental insights into virtually all aspects of cell behavior, including cell adhesion, migration, and differentiated function. However, cell responses to complex 3-D environments are difficult to adequately explore due to the large number of variables that must be controlled simultaneously. Here we describe an adaptable, automated approach for 3-D cell culture within hydrogel arrays. Our initial results demonstrate that the hydrogel network chemistry (both natural and synthetic), cell type, cell density, cell adhesion ligand density, and degradability within each array spot can be systematically varied to screen for environments that promote cell viability in a 3-D context. In a testbed application we then demonstrate that a hydrogel array format can be used to identify environments that promote viability of HL-1 cardiomyocytes, a cell line that has not been cultured previously in 3-D hydrogel matrices. Results demonstrate that the fibronectin-derived cell adhesion ligand RGDSP improves HL-1 viability in a dose-dependent manner, and that the effect of RGDSP is particularly pronounced in degrading hydrogel arrays. Importantly, in the presence of 70µM RGDSP, HL-1 cardiomyocyte viability does not decrease even after 7 days of culture in PEG hydrogels. Taken together, our results indicate that the adaptable, array-based format developed in this study may be useful as an enhanced throughput platform for 3-D culture of a variety of cell types.

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Section: Biological Characterization Of Hydrogel Arraysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An adaptable hydrogel array format for 3-dimensional cell culture and analysis

et al. 2008

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“…Such matrices include Matrigel (produced by EHS tumors, (Kleinman et al, 1986)), polymerized collagen (Grinnell et al, 2006;Grinnell et al, 1989), and other synthetic media (e.g., (Hwang et al, 2006)). Clearly, these matrices can regulate the growth of tumor cells in many ways, affecting their proliferation, morphology, survival signaling, invasive potential and response to chemotherapeutic agents (Griffith and Swartz, 2006).…”

Section: Introductionmentioning

confidence: 99%

Fibroblast-derived 3D matrix differentially regulates the growth and drug-responsiveness of human cancer cells

et al. 2008

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Recent studies have emphasized the importance of cellular microenvironment in modulating cell growth and signaling. In vitro, collagen matrices, Matrigel, and other synthetic support systems have been used to simulate in vivo microenvironments, and epithelial cells grown in these matrices manifest significant differences in proliferation, differentiation, response to drugs, and other parameters. However, these substrates do not closely resemble the mesenchymal microenvironment that is typically associated with advanced carcinomas in vivo, which is produced to a large extent by fibroblasts. In this study, we have evaluated the ability of a fibroblast-derived three-dimensional matrix to regulate the growth of a panel of 11 human tumor epithelial cell lines. Although proliferative and morphological responses to three-dimensional cues segregated independently, general responsiveness to the matrix correlated with the ability of matrix to influence drug responses. Fibroblast-derived three-dimensional matrix increased β1-integrin-dependent survival of a subset of human cancer cell lines during taxol treatment, while it sensitized or minimally influenced survival of other cells. β1-integrin-dependent changes in cell resistance to taxol did not correlate with degree of modulation of FAK and Akt, implying additional signaling factors are involved. Based on these results, we propose these matrices potentially have value as in vitro drug screening platforms.

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“…As the first step in chondrogenesis during the development of an embryo is the condensation of mesenchymal cells and a 3D tissue is required to repair cartilage defects, we have chosen 3D culture methods. More chondrogenic differentiation was observed in 3D cultures when compared to 2D cultures [98,111].…”

Section: Ebcell Ebsmentioning

confidence: 88%

“…When intact mouse EBs were encapsulated in alginate disks, and cultured either in basic proliferation medium, or medium supplemented with Dex, chondrogenesis was not enhanced, when compared with plated EBs [98]. However, when mouse EBs were encapsulated in a poly(ethylene glycol)-based hydrogel and cultured in serum-free chondrogenic medium containing TGFß1, a significant increase in cartilage markers was observed compared to 2D plating [111]. Addition of glucosamine to the chondrogenic medium resulted in improved cartilage matrix production [112].…”

Section: Cartilage Tissue Engineering Using Hydrogelsmentioning

confidence: 96%