Primary murine limb bud mesenchymal cells in long-term culture complete chondrocyte differentiation: TGF-β delays hypertrophy and PGE2 inhibits terminal differentiation

Zhang, Xinping; Ziran, Navid; Goater, J.Jeffery; Schwarz, Edward M.; Puzas, J. Edward; Rosier, Randy N.; Zuscik, Michael J.; Drissi, Hicham; O’Keefe, Regis J.

doi:10.1016/j.bone.2003.12.026

Cited by 107 publications

(118 citation statements)

References 48 publications

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“…Previous reports have shown micromass culture forwards to chondrocyte maturation and matrix calcification similar to in vivo development (Zhang et al, 2004, DeLise et al, 2000. These studies indicate that matrices of cartilage nodules formed in micromass culture could be similar to matrices of cartilage produced by mesenchymal cell in early chondrogenesis.…”

Section: Discussionsupporting

confidence: 57%

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Immunohistochemical Study of Amelogenin and Lysosome-Associate Membrane Proteins (LAMPs) in Cartilage

Hatakeyama

Oka

et al. 2014

Int. J. Morphol.

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SUMMARY:Amelogenin is one of the enamel matrix proteins secreted by ameloblasts during enamel formation in tooth development. Recent studies showed that the amelogenin is expressed in chondrocyte. Lysosome-associated membrane proteins (LAMPs) have been identified as binding partner proteins to amelogenin and it has been suggested they act as signaling receptors of amelogenin. The purpose of this study is to clarify the localization of amelogenin and LAMPs in growth plate cartilage and cartilaginous nodules in micromass culture. Mouse knee joints including tibia growth plate at 4 weeks old and micromass cultures of limb bud mesenchymal cells after 2 weeks were fixed in paraformaldehyde, routinely processed, sections were cut and immunostained with amelogenin, collagen type II and type X, LAMP-1 and -3. The positive immunoreaction of amelogenin was observed both in proliferation and hypertrophic zone cartilage of growth plate after enzymatic pretreatment in immunostaining. Furthermore, cartilaginous nodules in micromass culture were immunopositive to amelogenin. The chondrocytes in the proliferation zone of the growth plate were immunopositive to LAMP-1 but weakly stained in the chondrocytes of hypertrophic zone. These observations indicate that amelogenin may be present in cartilage matrix produced in vivo and in vitro and amelogenin may involve cartilage formation through the LAMP-1 signaling pathway.

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Section: Discussionsupporting

confidence: 57%

“…Micromass culture of mouse limb bud mesenchymal cells was performed as previously reported with minor modification (Hatakeyama et al, 2004, Zhang et al, 2004. Briefly, timed pregnant C57/BL6 mice were obtained and embryos were collected at embryonic day 10 (E10).…”

Section: Methodsmentioning

confidence: 99%

Immunohistochemical Study of Amelogenin and Lysosome-Associate Membrane Proteins (LAMPs) in Cartilage

Hatakeyama

Oka

et al. 2014

Int. J. Morphol.

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“…Isolated chondrocytes were maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% (v/v) fetal bovine serum, 50 g/ml streptomycin, and 50 units/ml penicillin. Mesenchymal cells were isolated from the limb buds of 11.5 dpc ICR mouse embryos and maintained as micromass culture to induce chondrogenesis and hypertrophic maturation (13,14). Briefly, 4.0 ϫ 10 7 cells/ml in Dulbecco's modified Eagle's medium/F-12 medium (2:3) containing 10% (v/v) fetal bovine serum, 50 g/ml streptomycin, and 50 units/ml penicillin were spotted as 15-l drops on culture dishes to induce chondrogenesis for 6 days.…”

Section: Methodsmentioning

confidence: 99%

“…Chondrogenesis was determined by examining the expression of collagen IIB and aggrecan by RT-PCR, as described below, or Alcian blue staining to detect accumulation of sulfated proteoglycans (15). Hypertrophic maturation of chondrocytes was determined by examining the expression of markers, such as collagen X and matrix metalloproteinase (MMP)-13, by reverse transcription-PCR (RT-PCR) or Alizarin Red staining to detect mineralization (13,14).…”

Section: Methodsmentioning

confidence: 99%

Cytokine-like 1 (CYTL1) Regulates the Chondrogenesis of Mesenchymal Cells

Kim

Ryoo

Chun

2007

Journal of Biological Chemistry

101

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To identify novel molecules regulating chondrogenesis and cartilage development, we screened a cartilage-specific expressed sequence tag data base. Cytokine-like 1 (Cytl1), a possible cytokine candidate with unknown function that was originally identified in bone marrow-derived CD34-positive cells, was selected for functional characterization. In view of the initial observation that Cytl1 is predominantly expressed in chondrocytes and cartilage, we investigated its possible role in chondrogenesis and hypertrophic maturation of chondrocytes. Cytl1 expression was very low in mesenchymal cells, dramatically increased during chondrogenesis, and decreased during hypertrophic maturation, both in vivo and in vitro. The role of Cytl1 in chondrogenesis and hypertrophic maturation was examined by treating chondrifying mesenchymal cells with exogenous Cytl1 or ectopic expression of Cytl1. Notably, exogenous Cytl1 caused chondrogenic differentiation of mouse limb bud mesenchymal cells during micromass culture. Lentivirus-mediated overexpression of Cytl1 additionally induced chondrogenic differentiation of mesenchymal cells. However, Cytl1 did not affect the hypertrophic maturation of chondrocytes. Cytl1 exerted its chondrogenic effect via stimulation of Sox9 transcriptional activity. In addition, Cytl1 caused expression of insulin-like growth factor 1, which has a capacity to induce chondrogenesis. Thus, our results collectively suggest that chondrocyte-specific Cytl1 regulates chondrogenesis as a novel autocrine factor, but not hypertrophic maturation of chondrocytes during cartilage development.Cartilage formation during embryonic development begins with the aggregation of mesenchymal cells, which ultimately differentiate into chondrocytes. Differentiated chondrocytes proliferate rapidly and secrete a cartilage-specific extracellular matrix such as type II collagen and sulfated proteoglycan to form cartilage. The cartilage serves as a template for endochondral ossification, which requires the maturation of hypertrophic chondrocytes (1-4). These sequential events during chondrogenesis and cartilage formation are precisely regulated by various growth factors released from cartilage elements and perichondrium. Secreted growth factors exert their effects by modulating intracellular signaling (1, 2). Although several regulatory growth factors have been identified, including bone morphogenetic proteins, fibroblast growth factors, insulin-like growth factor-1 (IGF-1), 2 transforming growth factor-␤, and parathyroid hormone-related peptide, the precise mechanisms of regulation of chondrogenesis and cartilage development remain to be elucidated. In this study, we analyze a cartilagespecific expressed sequence tag (EST) data base in an attempt to identify novel molecules that modulate chondrogenesis and cartilage development.The EST data base provides important information on novel genes displaying tissue-specific expression profiles (5-7). We analyzed the human cartilage UniGene library (8), and selected Cytl1 as a possible candid...

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“…In addition to its dual role in osteoblastogenesis, TGF-b signaling exerts a similar dual role in chondrogenesis by inducing the condensation of undifferentiated mesenchymal cells, stimulating their commitment to the chondrogenic lineage, and promoting proliferation of chondrocytes and deposition of cartilage-specific ECM, but also inhibiting terminal differentiation of chondrocytes (Leonard et al 1991;Ballock et al 1993;Mackay et al 1998;Worster et al 2000;Yang et al 2001;Tuli et al 2003;Zhang et al 2004;Song et al 2007;Mueller and Tuan 2008;Furumatsu et al 2009). BMP signaling, in contrast, is dispensable for the initial step of mesenchymal condensation but required for commitment to the chondrogenic lineage, proliferation, differentiation, and maturation of chondrocytes (Li et al 2003;Horiki et al 2004;Yoon et al 2005;Retting et al 2009;Culbert et al 2014).…”

Section: Regulation Of Chondrogenesis and Longitudinal Bone Growth Bymentioning

confidence: 99%

TGF-β Family Signaling in Connective Tissue and Skeletal Diseases

MacFarlane

Haupt

Dietz

et al. 2017

Cold Spring Harb Perspect Biol

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The transforming growth factor b (TGF-b) family of signaling molecules, which includes TGF-bs, activins, inhibins, and numerous bone morphogenetic proteins (BMPs) and growth and differentiation factors (GDFs), has important functions in all cells and tissues, including soft connective tissues and the skeleton. Specific TGF-b family members play different roles in these tissues, and their activities are often balanced with those of other TGF-b family members and by interactions with other signaling pathways. Perturbations in TGF-b family pathways are associated with numerous human diseases with prominent involvement of the skeletal and cardiovascular systems. This review focuses on the role of this family of signaling molecules in the pathologies of connective tissues that manifest in rare genetic syndromes (e.g., syndromic presentations of thoracic aortic aneurysm), as well as in more common disorders (e.g., osteoarthritis and osteoporosis). Many of these diseases are caused by or result in pathological alterations of the complex relationship between the TGF-b family of signaling mediators and the extracellular matrix in connective tissues.T he transforming growth factor b (TGF-b) family of cytokines comprises the three TGF-b proteins (TGF-b1, TGF-b2, and TGFb3) and related growth and differentiation factors such as activins, inhibins, bone morphogenic proteins (BMPs), and growth and differentiation factors (GDFs). These molecules play critical roles both in normal development and in several pathological conditions, including inflammation, fibrosis, and cancer (reviewed in Li and Flavell 2006;Gordon and Blobe 2008;Ikushima and Miyazono 2010). This review focuses on the role of these proteins in development and homeostasis of the skeleton and other connective tissues (summarized in Fig. 1) and on the diseases that ensue when these pathways are altered. Aberrant signaling in these pathways has been associated with common connective 6 These authors contributed equally to this work.

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Primary murine limb bud mesenchymal cells in long-term culture complete chondrocyte differentiation: TGF-β delays hypertrophy and PGE2 inhibits terminal differentiation

Cited by 107 publications

References 48 publications

Immunohistochemical Study of Amelogenin and Lysosome-Associate Membrane Proteins (LAMPs) in Cartilage

Immunohistochemical Study of Amelogenin and Lysosome-Associate Membrane Proteins (LAMPs) in Cartilage

Cytokine-like 1 (CYTL1) Regulates the Chondrogenesis of Mesenchymal Cells

TGF-β Family Signaling in Connective Tissue and Skeletal Diseases

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