Mineralization and metabolic response in serially passaged adult rat bone cells

Williams, Daniel C.; Boder, George B.; Toomey, Richard E.; Paul, Donald C.; Hillman, Charles C.; King, Kathleen L.; Frank, Richard M. Van; Johnston, C. Conrad

doi:10.1007/bf02408633

Cited by 119 publications

(37 citation statements)

References 39 publications

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“…Cell conditions and nodule formation were routinely checked by phase contrast microscopy. The presence of mineralized nodules was determined by staining with alizarin red (Wako Pure Chemical) as described previously (31).…”

Section: Alizarin Red Stainingmentioning

confidence: 99%

The in vitro osteogenetic characteristics of primary osteoblastic cells from a rabbit calvarium

et al. 2008

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Previously, we showed that recombinant human bone morphogenetic protein-2 (rhBMP-2) increased bone augmentation beyond the skeletal envelope within a titanium cap in a rabbit calvarium; many cuboidal osteoblastic cells were observed histologically. These results suggested that the new osteoblastic cells might have differentiated and matured via stimulation by rhBMP-2. To date, however, no studies have reported the characteristics of osteoblastic cells derived from adult rabbit calvarium, after addition of rhBMP-2. To determine the effects of rhBMP-2 on osteoblastic cells, we observed morphological characteristics and alkaline phosphatase activity of osteoblastic cells from an adult rabbit calvarium. The expression of proteins in the BMP signaling pathway and extracellular matrix were analyzed, and mineralized nodule formation was assessed. The alkaline phosphatase activity increased significantly after rhBMP-2 stimulation. The protein levels of phosphorylated-Smad1, Runx2, osteocalcin, osteopontin, and type I collagen were augmented by rhBMP-2 stimulation using Western blotting or ELISA; rhBMP-2 also stimulated mineralized nodule formation with alizarin red staining. The results suggest that primary osteoblastic cells derived from a rabbit calvarium have osteogenetic characteristics in vitro, underscoring the potential use of these cells as a model for studying bone formation. These cells may play an important role in in vivo bone augmentation in a rabbit experimental model. (J. Oral Sci. 50, [427][428][429][430][431][432][433][434] 2008)

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Section: Alizarin Red Stainingmentioning

confidence: 99%

The in vitro osteogenetic characteristics of primary osteoblastic cells from a rabbit calvarium

et al. 2008

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“…Cultures were fixed in 10% v/v formol saline for 5 min. Bone nodules were detected using a calcium specific dye Alizarin Red (pH 7.4) (27). Bone nodules per well were counted macroscopically.…”

Section: Genotyping Of Enos Gene-deficient Mice and Wild-type Controlsmentioning

confidence: 99%

Nitric Oxide Mediates 17β-Estradiol-Stimulated Human and Rodent Osteoblast Proliferation and Differentiation

O'Shaughnessy

Polak

Afzal

et al. 2000

Biochemical and Biophysical Research Communications

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“…However, the potential for division of such cells remains limited, and this precludes molecular studies. In addition, phenotypic changes with loss of osteoblastic properties often occurs in culture (47).…”

mentioning

confidence: 99%

An immortalized osteogenic cell line derived from mouse teratocarcinoma is able to mineralize in vivo and in vitro.

Kellermann

Buc-Caron

Marie

et al. 1990

The Journal of Cell Biology

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Abstract. The hybrid plasmid pK4 containing the early genes of the simian virus SV-40, under the control of the adenovirus type 5 Ela promoter, was introduced into the multipotent embryonal carcinoma (EC) 1003. Expression of the SV-40 oncogenes was observed at the EC cell stage, and this allowed the derivation of immortalized cells corresponding to early stages of differentiation.Among the immortalized mesodermal derivatives obtained, one clone, C1, is committed to the osteogenic pathway. C1 cells have a stable phenotype, synthesize type I collagen, and express alkaline phosphatase activity. Although immortalized and expressing the SV-40 T antigen, the cells continue to be able to differentiate in vivo and in vitro. In vivo, after injection into syngeneic mice, they produce osteosarcomas. In vitro, the cells form nodules and deposit a collagenous matrix that mineralizes, going to hydroxyapatite crystal formation, in the presence of/~-glycerophosphate. This clonal cell line, which originates from an embryonal carcinoma, therefore differentiates into osteogenic cells in vivo and in vitro.This immortalized cell line will be useful in identifying specific molecular markers of the osteogenic pathway, to investigate gene regulation during osteogenesis and to study the ontogeny of osteoblasts.N~ current view of the development of bone cells is that osteocytes derive from primitive mesoblastic cell precursors through a cascade of events. These involve, in the case of intramembranous ossification, (a) proliferation of primitive mesoblasts, (b) differentiation of these cells into an osteoprogenitor cell, then into osteoblasts, and (c) maturation of osteoblasts with synthesis of a collagen matrix and mineralization (5,10,39,44). Up until now, models for following bone differentiation in vitro have been of two types: (a) osteoblast-like clones explanted from normal bone, and (b) clones derived from osteogenic osteosarcomas.(a) The former are normal explanted cells capable of progressively synthesizing a bone-like tissue in the presence of organic phosphate and ascorbic acid (1,3,8,9,28,35,46). This process and its hormonal regulation can therefore be studied in vitro. Collagen I maturation, in particular, has been extensively studied (11). However, the potential for division of such cells remains limited, and this precludes molecular studies. In addition, phenotypic changes with loss of osteoblastic properties often occurs in culture (47).(b) Clones derived from rat osteosarcomas have also been useful in investigating the effects of hormones (parathyroid hormone; prostaglandins) and vitamins (retinoids and vitamin D3) on bone development (17,25,29,30,41). The formation of a mineralized matrix has, however, been shown to require either the implantation of the tumoral cells in diffusion chambers within the animal (43), or the growth of the ceils in agar (37).In both model systems, the major problems encountered are the heterogeneity of the cell population within a clone, and the lack of specific markers allowing formal identi...

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Mineralization and metabolic response in serially passaged adult rat bone cells

Cited by 119 publications

References 39 publications

The in vitro osteogenetic characteristics of primary osteoblastic cells from a rabbit calvarium

The in vitro osteogenetic characteristics of primary osteoblastic cells from a rabbit calvarium

Nitric Oxide Mediates 17β-Estradiol-Stimulated Human and Rodent Osteoblast Proliferation and Differentiation

An immortalized osteogenic cell line derived from mouse teratocarcinoma is able to mineralize in vivo and in vitro.

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