A Quadripotential Mesenchymal Progenitor Cell Isolated from the Marrow of an Adult Mouse

Dennis, James E.; Merriam, Anita P.; Awadallah, Amad; Yoo, Jung U.; Johnstone, Brian; Caplan, Arnold I.

doi:10.1359/jbmr.1999.14.5.700

Cited by 387 publications

(232 citation statements)

References 65 publications

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“…Mesenchymal stem cells have been used in multiple tissue-engineering applications, and they differentiate into multiple lineages, including osteoblasts, chondrocytes, adipocytes, and muscle/tendon forming cells [9][10][11]23]. The multipotent, immortalized BMSC line used here is primarily osteogenic [24] and has been used in models of spinal fusion [19] and in the femur [22].…”

Section: Discussionmentioning

confidence: 99%

The Effects of GDF-5 and Uniaxial Strain on Mesenchymal Stem Cells in 3-D Culture

Farng

Urdaneta²,

Barba³

et al. 2008

Clin Orthop Relat Res

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Recent endeavors in tissue engineering have attempted to identify the optimal parameters to create an artificial ligament. Both mechanical and biochemical stimulation have been used by others to independently modulate growth and differentiation, although few studies have explored their interactions. We applied previously described fabrication techniques to create a highly porous (90%-95% porosity, 212-300 lm), 3-D, bioabsorbable polymer scaffold (polycaprolactone). Scaffolds were coated with bovine collagen, and growth and differentiation factor 5 (GDF-5) was added to half of the scaffolds. Scaffolds were seeded with mesenchymal stem cells and cultured in a custom bioreactor under static or cyclic strain (10% strain, 0.33 Hz) conditions. After 48 hours, both mechanical stimulation and GDF-5 increased mRNA production of collagen I, II, and scleraxis compared to control; tenascin C production was not increased. Combining stimuli did not change gene expression; however, cellular metabolism was 1.7 times higher in scaffolds treated with both stimuli. We successfully grew a line of mesenchymal stem cells in 3-D culture, and our initial data indicate mechanical stimulation and GDF-5 influenced cellular activity and mRNA production; we did not, however, observe additive synergism with the mechanical and biological stimuli.

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Section: Discussionmentioning

confidence: 99%

The Effects of GDF-5 and Uniaxial Strain on Mesenchymal Stem Cells in 3-D Culture

Farng

Urdaneta²,

Barba³

et al. 2008

Clin Orthop Relat Res

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“…To this end, we used mouse bone marrow stromal cells, which are capable of differentiating into several different lineages, including osteoblasts, when cultured under the appropriate conditions (50,51). These bone marrow stromal cells were allowed to differentiate at confluence in the presence of ␤-glycerophosphate and L-ascorbate- 2-phosphate.…”

Section: Trps1 Expression and Ose2 Dna Binding In Differentiating Bonementioning

confidence: 99%

Identification of the GATA Factor TRPS1 as a Repressor of the Osteocalcin Promoter

Piscopo

Johansen

Derynck

2009

Journal of Biological Chemistry

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A proteomic analysis of proteins bound to the osteocalcin OSE2 sequence of the mouse osteocalcin promoter identified TRPS1 as a regulator of osteocalcin transcription. Mutations in the TRPS1 gene are responsible for human tricho-rhino-phalangeal syndrome, which is characterized by skeletal and craniofacial abnormalities. TRPS1 has been shown to bind regulatory promoter sequences containing GATA consensus binding sites and to repress transcription of genes involved in chondrocyte differentiation. Here we show that TRPS1 can directly bind the osteocalcin promoter in the presence or absence of Runx2. TRPS1 binds through a GATA binding sequence in the proximal promoter of the osteocalcin gene. The GATA binding site is conserved in mice, humans, and rats, although its location and orientation are not. Mutation of the mouse or human GATA binding sequence abrogates binding of TRPS1 to the osteocalcin promoter. We show that TRPS1 is expressed in osteosarcoma cells and upon induction of osteoblast differentiation in primary mouse bone marrow stromal cells and that TRPS1 regulates the expression of osteocalcin in both cell types. The expression of TRPS1 modulates mineralized bone matrix formation in differentiating osteoblast cells. These data suggest a role for TRPS1 in osteoblast differentiation, in addition to its previously described role in chondrogenesis.Runx2, previously described as CBFA1, OSF2, AML3, or PEPB2␣A, is a runt homology domain transcription factor and plays a key role in driving osteoblast differentiation. Runx2 expression, which occurs in differentiating and mature osteoblasts, is essential for bone formation, as illustrated by the phenotype of Runx2-deficient mice, which lack osteoblasts and have skeletons that are primarily composed of immature chondrocytes (1-3). Haploinsufficiency for RUNX2 in humans results in cleidocranial dysplasia, a syndrome that is characterized by delayed endochondral and intramembranous ossification (4). Runx2 transactivates genes involved in the deposition of bone matrix, e.g. those encoding osteocalcin, collagen IA1, osteopontin, and matrix metalloproteinase 13 (MMP13), and regulation of osteoclastogenesis, e.g. the genes encoding RankL and osteoprotegerin (reviewed in Ref. 5). Although Runx2 is expressed in cells of the osteoblast lineage throughout skeletal development, its levels increase only 1.5-2-fold as preosteoblasts differentiate into mature osteoblasts. Furthermore, the amount of Runx2 bound to target promoters, as measured by chromatin immunoprecipitation, is relatively unchanged throughout differentiation (6), yet the targets it regulates have distinct patterns of expression. These observations suggest that the regulation of Runx2 activity, either by post-translational modification or by the binding of co-regulators, plays a substantial role in defining the Runx2-mediated activation of target genes. Various proteins have been shown to interact with Runx2 and modulate its activity, including Smad3, CREB 2 -binding protein (CBP) HES1, Groucho/TLE, and histone d...

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“…[1][2][3] Induction of osteoprogenitor differentiation of MSCs by dexamethasone (Dex) is well known, 4,5 and culture supplementation with Dex at 10 À8 M has been reported to promote osteoprogenitor cell differentiation in marrow stromal cells. 6,7 In addition, Dex is also a component of medium used for differentiation into chondroblasts, myocytes, and adipocytes in vitro.…”

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confidence: 99%

Dexamethasone inhibition of confluence‐induced apoptosis in human mesenchymal stem cells

Song

Caplan

Dennis

2008

Journal Orthopaedic Research

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Human mesenchymal stem cells (MSCs) have been extensively characterized with respect to their in vitro expansion and differentiation potential, especially with respect to osteogenesis. Dexamethasone (Dex) is a well-known inducer of osteogenic differentiation of MSCs, but little is known about the effect of Dex treatment on apoptosis in MSCs. In this study, apoptosis in MSCs was examined with respect to cell density and Dex supplementation, using DAPI staining and DNA fragmentation ELISA Assay. In MSC cultures initiated at 1.0, 3.0, and 9.0 Â 10 3 cells per cm 2 , it was found that higher MSC density correlated with increased apoptosis and that this apoptotic effect was diminished in cultures containing 100 nM Dex. MSCs and fibroblasts were co-cultured, along with empty insert controls, and assayed for apoptosis by ELISA and DAPI counts to determine if soluble factors accounted for the cell density-related apoptosis. No difference was seen between MSCs cultured with inserts containing either MSCs, fibroblasts, or empty control. To determine cell contact effects, BrdU-labeled MSCs were cultured alone or with unlabeled chondrocytes at 2Â and 8Â the number of MSCs, with and without Dex, and apoptosis levels quantified. The results showed increased apoptosis at greater cell densities, and that the amount of apoptosis was greatly diminished in cultures containing Dex. These results show that apoptosis in MSCs is cell density-related, requires direct cell contact, and that Dex treatment reduces or eliminates this density-related apoptosis. These results may impact how MSCs should be cultured for clinical applications. Keywords: dexamethasone; mesenchymal stem cells; apoptosis Human bone marrow-derived mesenchymal stem cells (MSCs) are a rare population of undifferentiated cells that have the capacity to self-renew and can differentiate into mesodermal phenotypes including osteoblasts, chondrocytes, myocytes, and adipocytes. 1-3 Induction of osteoprogenitor differentiation of MSCs by dexamethasone (Dex) is well known, 4,5 and culture supplementation with Dex at 10 À8 M has been reported to promote osteoprogenitor cell differentiation in marrow stromal cells. 6,7 In addition, Dex is also a component of medium used for differentiation into chondroblasts, myocytes, and adipocytes in vitro. [8][9][10] With respect to Dex effects on apoptosis, the results have often been contradictory, depending upon the cell type being studied. For example, Dex treatment has been shown to induce apoptosis in thymocytes, 11,12 lymphocytes, 13-15 some tumor cells, 16,17 and respiratory epithelium, 18,19 whereas primary cultured hepatocytes, 20 neutrophils, 21,22 and some solid tumors 23,24 show reduced apoptosis after Dex treatment.Interestingly, while Dex has been used extensively with MSCs as an inductive factor for multiple end-stage phenotypes, few studies have assessed the effects of Dex on proliferation and apoptosis. To date, only one other study addressed the issue of Dex effects on MSC apoptosis wherein it was shown that hu...

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A Quadripotential Mesenchymal Progenitor Cell Isolated from the Marrow of an Adult Mouse

Cited by 387 publications

References 65 publications

The Effects of GDF-5 and Uniaxial Strain on Mesenchymal Stem Cells in 3-D Culture

The Effects of GDF-5 and Uniaxial Strain on Mesenchymal Stem Cells in 3-D Culture

Identification of the GATA Factor TRPS1 as a Repressor of the Osteocalcin Promoter

Dexamethasone inhibition of confluence‐induced apoptosis in human mesenchymal stem cells

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