Calvaria Derived Osteogenic Cells: Phenotypic Expression in Culture

Binderman, Itzhak; Berger, Esther; Fine, N.; Shimshoni, Zvi; Harell, A; Sömjen, Dalia

doi:10.3109/03008208909023873

Cited by 13 publications

(9 citation statements)

References 12 publications

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“…Both CK activity and DNA synthesis were stimulated by all of the cell fragments tested except hPTH-(34-47) ( [18].…”

Section: Resultsmentioning

confidence: 98%

“…In the present paper, we used two osteoblast-like cell cultures and one chondroblast-like cell culture of rat cells to demonstrate that the same functional domain of the PTH molecule which is active in an avian system is responsible for the mitogenic response in the rat. For this work, we used two assay systems: (1) the incorporation of [3H]thymidine, a measure of DNA synthesis, and (2) the stimulation of the specific activity of the brain-type isoenzyme of creatine kinase (CK [17][18][19]), the enzyme that regulates intracellular levels of ATP and ADP [20]. Whereas [3H]thymidine incorporation is a relatively late response to a mitogen, the increased activity of CK reflects a very early response to mitotic stimulation, and its rapid and sensitive assay makes CK a convenient enzyme marker for hormone action [12,17,19,21].…”

Section: Introductionmentioning

confidence: 99%

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Stimulation by defined parathyroid hormone fragments of cell proliferation in skeletal-derived cell cultures

Sömjen¹,

Binderman

Schlüter

et al. 1990

Biochemical Journal

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We have reported previously that parathyroid hormone (PTH) acts on cultured bone cells to stimulate creatine kinase (CK) activity and [3H]thymidine incorporation into DNA via phosphoinositide turnover, in addition to its other actions via increased cyclic AMP production. We also found that mid-region fragments of PTH stimulate [3H]thymidine incorporation into avian chondrocytes. In the present study of mammalian systems, we demonstrate differential effects of defined synthetic PTH fragments on CK activity and DNA synthesis, as compared with cyclic AMP production, in osteoblast-enriched embryonic rat calvaria cell cultures, in an osteoblast-like clone of rat osteosarcoma cells (ROS 17/2.8) and in chondroblasts from rat epiphysial cartilage cell cultures. Unlike full-length bovine (b)PTH-(1-84) or the fully effective shorter fragment human (h)PTH-(1-34), fragments lacking the N-terminal region of the hormone did not increase cyclic AMP formation, whereas they did stimulate increases in both DNA synthesis and CK activity. Moreover, the PTH fragment hPTH-(28-48) at 10 /M inhibited the increase in cyclic AMP caused by 10 nm-bPTH-(1-84). The increase of CK activity in ROS 17/2.8 cells caused by bPTH-(1-84) or hPTH-(28-48) was completely inhibited by either cycloheximide or actinomycin D, as was shown previously for rat calvaria cell cultures. These results indicated the presence of a functional domain of PTH in the central part of the molecule which exerts its mitogenic-related effects on osteoblast-and chondroblast-like cells in a cyclic AMP-independent manner. Since cyclic AMP formation by PTH leads to bone resorption, specific mid-region fragments of PTH might prove suitable for use in vivo to induce bone formation without concomitant resorption. INTRODUCTIONIn conjunction with its regulation of calcium homeostasis, parathyroid hormone (PTH) can stimulate both net bone formation and resorption, depending on the conditions of application. Increased levels of cyclic AMP [1] are considered to mediate the action of PTH in causing bone resorption [2] and stimulation of ornithine decarboxylase activity [3,4]. On the other hand, PTH has also been shown to exert proliferative and anabolic effects,

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“…Both CK activity and DNA synthesis were stimulated by all of the cell fragments tested except hPTH-(34-47) ( [18].…”

Section: Resultsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Stimulation by defined parathyroid hormone fragments of cell proliferation in skeletal-derived cell cultures

Sömjen¹,

Binderman

Schlüter

et al. 1990

Biochemical Journal

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“…In brief, samples of the trabecular surface of the iliac crest or long bones were cut into 1 mm 3 pieces and extensively and repeatedly washed with phosphate buffered saline (PBS) to remove blood components. The explants, with no enzymatic digestion, were seeded in 100 mm diameter tissue culture dishes and incubated in DMEM medium without Ca 2+ , to avoid fibroblastic growth [25,26], containing 10% fetal calf serum (FCS) and antibiotics. Cell outgrowth from the bone explants was apparent after 6-10 days.…”

Section: Cell Culturesmentioning

confidence: 99%

Treatment with non-hypercalcemic analogs of 1,25-dihydroxyvitamin D3 increases responsiveness to 17β-estradiol, dihydrotestosterone or raloxifene in primary human osteoblasts

Katzburg

Hendel

Waisman

et al. 2004

The Journal of Steroid Biochemistry and Molecular Biology

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“…Furthermore, it could also be very useful for characterisation of osteocytes in bone cell culture systems. Isolated primary cultures from rat calvaria (Ecarot-Charrier et al 1983;Binderman et al 1989) and the rat osteosarcoma cell line ROS 17/2.8 (Majeska et al 1980) are well-established bone cell cultures which are often used for in vitro studies of osteogenesis. Up to now, differentiation markers for cultured osteoblastic cells have been restricted to alkaline phosphatase (Majeska and Rodan 1982;Herbert et al 1997), osteonectin, osteocalcin, bone sialoprotein and osteopontin (Collin et al 1992;Nanci et al 1996;Moursi et al 1997), all of which occur over a considerably large span of the osteoblastic-osteocytic differentiation period (Bruder et al 1997).…”

Section: Fig 5a-fmentioning

confidence: 99%

Immunohistochemical investigations on the differentiation marker protein E11 in rat calvaria, calvaria cell culture and the osteoblastic cell line ROS 17/2.8

Schulze

Witt

Kasper

et al. 1999

Histochemistry and Cell Biology

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Until now, many extracellular matrix proteins, e.g. osteopontin and osteonectin, have been used to determine a cell's osteogenic maturation. The disadvantage in evaluation of these proteins is their relative wide-ranging appearance throughout the osteogenic differentiation process. Thus, the aim of this study was to establish an immunohistochemical setup using E11, a marker that binds selectively to cells of the late osteogenic cell lineage. In addition, the histochemical expression of the bone matrix proteins osteonectin, osteopontin and fibronectin was compared to that of E11 using monoclonal antibodies. For light microscopical detection of osteogenic markers in cultured cells we developed a simple paraffin technique using a fibrin glue as embedding medium. This allows the handling of cultured cells such as a tissue sample and includes the use of stored biological specimens for further immunohistochemical experiments. We used newborn rat calvariae for whole tissue preparations and for isolation and cultivation of bone cells. In addition, we included the rat osteosarcoma cell line ROS 17/2.8 in this study. For the first time, we have localised E11 in osteocytes of rat calvaria preparations at the electron microscopical level. E11 was detected at plasma membranes of osteocytes and their processes, but not at those of osteoblasts. Accompanying experiments with cultured newborn rat calvaria cells and ROS 17/2.8 cells revealed E11 reactivity on a subset of cells. The results obtained confirm the suitability of the differentiation marker E11 as a sensitive instrument for the characterisation of bone cell culture systems.

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Calvaria Derived Osteogenic Cells: Phenotypic Expression in Culture

Cited by 13 publications

References 12 publications

Stimulation by defined parathyroid hormone fragments of cell proliferation in skeletal-derived cell cultures

Stimulation by defined parathyroid hormone fragments of cell proliferation in skeletal-derived cell cultures

Treatment with non-hypercalcemic analogs of 1,25-dihydroxyvitamin D3 increases responsiveness to 17β-estradiol, dihydrotestosterone or raloxifene in primary human osteoblasts

Immunohistochemical investigations on the differentiation marker protein E11 in rat calvaria, calvaria cell culture and the osteoblastic cell line ROS 17/2.8

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