Osteoprogenitor cells in cell populations derived from mouse and rat calvaria differ in their response to corticosterone, cortisol, and cortisone1

Bellows, C. G.; Ciaccia, Antonio; Heersche, J. N. M.

doi:10.1016/s8756-3282(98)00084-2

Cited by 55 publications

(33 citation statements)

References 28 publications

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“…Primary Cultures of Osteoblast-enriched Cells-Primary cultures of osteoblast-enriched cells were prepared as described previously (25). Briefly, mouse osteoblast-enriched cells were prepared from the calvariae of 3-to 5-day old newborn mice as follows.…”

Section: Methodsmentioning

confidence: 99%

ANA Deficiency Enhances Bone Morphogenetic Protein-induced Ectopic Bone Formation via Transcriptional Events

Miyai

Yoneda

Hasegawa

et al. 2009

Journal of Biological Chemistry

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Ectopic bone formation after joint replacement or brain injury in humans is a serious complication that causes immobility of joints and severe pain. However, mechanisms underlying such ectopic bone formation are not fully understood. Bone morphogenetic protein (BMPs) are defined as inducers of ectopic bone formation, and they are regulated by several types of inhibitors. ANA is an antiproliferative molecule that belongs to Tob/BTG family, but its activity in bone metabolism has not been known. Here, we examined the role of ANA on ectopic bone formation activity of BMP. In ANA-deficient and wild-type mice, BMP2 was implanted to induce ectopic bone formation in muscle. ANA deficiency increased mass of newly formed bone in vivo compared with wild-type based on 3D-CT analyses. ANA mRNA was expressed in bone in vivo as well as in osteoblastic cells in vitro. Such ANA mRNA levels were increased by BMP2 treatment in MC3T3-E1 osteoblastic cells. Overexpression of ANA suppressed BMP-induced expression of luciferase reporter gene linked to BMP response elements in these cells. Conversely, ANA mRNA knockdown by small interference RNA enhanced the BMP-dependent BMP response element reporter expression. It also enhanced BMP-induced osteoblastic differentiation in muscle-derived C2C12 cells. Immunoprecipitation assay indicated that ANA interacts with Smad8. Thus, ANA is a suppressor of ectopic bone formation induced by BMP, and this inhibitory ANA activity is a part of the negative feedback regulation of BMP function.Ectopic or heterotopic bone formation is one of the major complications associated with joint replacement surgery and brain injury (1, 2). Such pathological ectopic bone formation and heterotopic ossification cause ankylosis of joints or persistent pain and significantly deteriorated quality of life. However, the molecules involved in these ectopic bone formation events have not been fully understood.Bone morphogenetic proteins (BMPs) 3 were first identified as an inducer of ectopic bone formation (3, 4). Although BMP2 is required for fracture healing, skeletal development of the limbs was normal in the absence of BMP2 (5). In contrast, BMP is absolutely required for ectopic bone formation in muscle (3), and no other growth factors could replace such potent ectopic bone formation activity of this molecule. Thus, BMP action on ectopic bone formation is one of the unique features of this molecule.BMP activity is under the control of several groups of modulators against its signaling pathway. One of these groups of modulators that control BMP2 signaling in osteoblasts includes the Tob/BTG antiproliferative protein family, comprising Btg1, Btg2, Tob, Tob2, Pc3, and ANA. These molecules share a homologous region (BTG/Tob homology domain) in their N-terminal ends (6) and suppress cell proliferation when they are overexpressed in NIH3T3 cells (7-13).Among the members of Tob/BTG family, Tob was shown to be a BMP antagonist previously (14, 15). However, effects of other members of Tob/BTG family on BMP activity have not y...

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Section: Methodsmentioning

confidence: 99%

ANA Deficiency Enhances Bone Morphogenetic Protein-induced Ectopic Bone Formation via Transcriptional Events

Miyai

Yoneda

Hasegawa

et al. 2009

Journal of Biological Chemistry

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“…6,11,13) The presence of mineralized bone nodules in FRC cultures was demonstrated with von Kossa staining. 14) On day 14, triplicate cultures of each fraction of FRC cells in 24-well plates were rinsed once with PBS and fixed with 100% ethanol for 10 min.…”

Section: Culture Of Frc Cellsmentioning

confidence: 99%

“…In addition, these proteins are expressed in a temporal pattern similar to what is seen in vivo. 6,[8][9][10][11] Nevertheless, it is still unknown which fraction of cells obtained from sequential digestions of FRC exhibits unique osteoblastic phenotypes and is suitable for analyses of phenotypic changes in cultures during the differentiation process, since pooled fractions of FRC cells have been frequently used.In the present study, we fractionated FRC cells by sequential enzymatic digestions and characterized the osteoblastic differentiation ability of each fraction of cells. Thereafter, using a rich fraction of osteoprogenitor cells, the production of bone matrix proteins, matrix maturation manifested by cross-linking, and gene expression were investigated for analyses of changes in osteoblastic phenotype.…”

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confidence: 99%

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confidence: 99%

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Progressive Development of the Osteoblast Phenotype during Differentiation of Osteoprogenitor Cells Derived from Fetal Rat Calvaria: Model for in Vitro Bone Formation.

Yamamoto

Furuya

Hanada

2002

Biological & Pharmaceutical Bulletin

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Bone consists of various types of cells and an abundant mineralized extracellular matrix. Bone formation as well as bone resorption are essential processes for the maintenance of normal bone structure and calcium homeostasis in the body. In addition to its physiological role, bone formation plays important roles in pathological disorders such as osteoporosis, metabolic bone diseases and bone fracture. Osteoblasts are the primary cells responsible for bone formation, and are believed to originate form mesenchymal osteoprogenitor cells within skeletal tissues.1,2) Therefore, the elucidation of the function and differentiation process of the osteoprogenitor cells will allow us to understand a major portion of the process of bone metabolism.Many model systems have been used to examine bone cell metabolism; these include cultures of calvariae and long bone explants, continuously cultured osteosarcoma cell lines, as well as primary cultures of osteoblastic cells.3,4) Each model system offers unique advantages to increase our understanding of bone development and metabolism. Of these, clonal osteosarcoma cell lines are thought to be useful tools because of their relatively homogeneous population of osteoblastic cells. However, since osteosarcoma cell lines express intrinsic properties reflecting specific stages of osteoblast differentiation, only a limited number of cell lines retain the capacity to form mineralized bones in vitro, reflecting terminal differentiation of osteoblasts. Another useful culture system is primary osteoblastic cells isolated from bones by enzymatic digestion, which have various phenotypic characteristics of osteoblasts including the potential to form mineralized bones in vitro. However, the spatial and temporal expression patterns of osteoblast phenotypes in this system have not been fully understood, since primary cultures of osteobastic cells consist of a heterogeneous cell population and these cultures often lose bone formation ability after several passages of cells. 5,6) Osteoblast precursor cells isolated by the enzymatic digestion of fetal rat calvaria (FRC) can be grown in culture to develop into mature osteoblasts which form mineralized bone nodules. [6][7][8] This process can be accelerated by a treatment with glucocorticoid hormones such as dexamethasone (Dex). Dex-stimulated cultures of FRC cells have been extensively characterized and have been shown to produce high levels of alkaline phosphatase (ALP), type I collagen, osteonectin, osteocalcin, and other noncollagenous proteins that are found in bone. In addition, these proteins are expressed in a temporal pattern similar to what is seen in vivo. 6,[8][9][10][11] Nevertheless, it is still unknown which fraction of cells obtained from sequential digestions of FRC exhibits unique osteoblastic phenotypes and is suitable for analyses of phenotypic changes in cultures during the differentiation process, since pooled fractions of FRC cells have been frequently used.In the present study, we fractionated FRC cells by sequential enzymatic...

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“…Ascorbic acid (50 g͞ml) was added at confluency, and ␤-glycerophosphate (10 mM) was added after nodules had formed (Ϸday 10 after confluency). Murine calvarial cells from day 3 neonatal mice were prepared by enzymatic digestion as described (27) and cultured in ␣MEM supplemented with 10% FCS and antibiotics. For in vitro osteogenic differentiation, mouse calvarial cells were cultured until confluency, followed by addition of dexamethasone (10 nM) and ascorbic acid (50 g͞ml).…”

Section: Methodsmentioning

confidence: 99%

The immune regulatory protein B7-H3 promotes osteoblast differentiation and bone mineralization

Suh

Wang

Jheon

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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B7-H3, a member of the B7 family of the Ig superfamily proteins, is expressed on the surface of the antigen-presenting cells and down-regulates T cell functions by engaging an unknown counterreceptor on T cells. Although B7-H3 is ubiquitously expressed, its potential nonimmune functions have not been addressed. We found that B7-H3 is highly expressed in developing bones during embryogenesis and that its expression increases as osteoblast precursor cells differentiate into mature osteoblasts. In vitro bone formation by osteoblastic cells was inhibited when B7-H3 function was interrupted by the soluble recombinant protein B7-H3-Fc. Analysis of calvarial cells derived from neonatal B7-H3 knockout (KO) mice revealed normal numbers of osteoblast precursor cells possessing a normal proliferative capacity. However, the B7-H3-deficient calvarial cells exhibited impaired osteogenic differentiation, resulting in decreased mineralized bone formation in vitro. These results suggest that B7-H3 is required for the later phase of osteoblast differentiation. Although B7-H3 KO mice had no gross skeletal abnormalities, they displayed a lower bone mineral density in cortical (but not trabecular) bones compared with WT controls. Consistent with the reduced bone mineral density, the femurs of B7-H3 KO mice were more susceptible to bone fracture compared with those of WT mice. Taken together, these results indicate that B7-H3 and its unknown counterreceptor play a positive regulatory role in bone formation. In addition, our findings identified B7-H3 as another molecule that has a dual role in the bone-immune interface.

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Osteoprogenitor cells in cell populations derived from mouse and rat calvaria differ in their response to corticosterone, cortisol, and cortisone1

Cited by 55 publications

References 28 publications

ANA Deficiency Enhances Bone Morphogenetic Protein-induced Ectopic Bone Formation via Transcriptional Events

ANA Deficiency Enhances Bone Morphogenetic Protein-induced Ectopic Bone Formation via Transcriptional Events

Progressive Development of the Osteoblast Phenotype during Differentiation of Osteoprogenitor Cells Derived from Fetal Rat Calvaria: Model for in Vitro Bone Formation.

The immune regulatory protein B7-H3 promotes osteoblast differentiation and bone mineralization

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