Establishment of an osteogenic cell line derived from adult mouse bone marrow stroma by use of a recombinant retrovirus

Mathieu, Éric; Schoeters, Greet; Plaetse, F. Vander; Merregaert, J.

doi:10.1007/bf00301635

Cited by 27 publications

(9 citation statements)

References 38 publications

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“…Despite the small number of cells, the retroviral transfection was extremely effective in establishing stably transfected cell lines. As reported previously, this methodology has 100‐fold or higher stable transfection rates compared with more conventional methods, such as calcium chloride precipitation or electroporation (39,42,51‐53,54) . Thus, a large number of cell lines were available for characterization.…”

Section: Discussionmentioning

confidence: 85%

Development and Characterization of Conditionally Immortalized Osteoblast Precursor Cell Lines from Human Bone Marrow Stroma

Hicok

Thomas

Gori

et al. 1998

Journal of Bone and Mineral Research

116

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Although the differentiation of mature osteoblasts has been well studied, there is still a need for a convenient way to study preosteoblast differentiation. Our laboratory has recently described a method for isolating small numbers of authentic osteoblast precursor cells from human bone marrow (Rickard et al., J Bone Miner Res 11:312-324, 1996). Here we describe the conditional immortalization of these cells by retroviral transfection with the amphotrophic vector, pZipSV40tsa58, which encodes for a temperature-sensitive mutant form of the simian virus large T-antigen. At the permissive temperature of 34°C, the cell lines proliferated, but differentiation was arrested, whereas at the restrictive temperature of 39.5°C, proliferation was decreased and differentiation was induced. As assessed by semiquantitative reverse transcriptase PCR after 4 days of culture at 39.5°C, the six cell lines expressed similar mRNA levels both constitutively and in response to dexamethasone (

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Section: Discussionmentioning

confidence: 85%

Development and Characterization of Conditionally Immortalized Osteoblast Precursor Cell Lines from Human Bone Marrow Stroma

Hicok

Thomas

Gori

et al. 1998

Journal of Bone and Mineral Research

116

View full text Add to dashboard Cite

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“…1990) or from cloned cell lines derived from marrow stroma (Benayahu et al 1989, Mathieu et al 1991 .…”

Section: Introductionmentioning

confidence: 99%

High Radiosensitivity of the Mineralization Capacity of Adult Murine Bone Marrowin Vitroto Continuous α-irradiation Compared to Acute X-irradiation

Schoeters

Plaetse

Heuvel

1992

International Journal of Radiation Biology

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Adult BALB/c mice, injected with osteosarcomogenic amounts of 241Am (between 40 and 500 Bq/g mouse) showed an impaired mineralization capacity of their femoral bone marrow . This effect persisted until at least 1 year after "'Am injection and was expressed after incubation of bone marrow cells in vitro in conditions allowing osteogenic differentiation. The mineralization capacity of marrow in vitro was evaluated by measurement of "Sr uptake from the tissue culture medium . Two osteogenic assays were used : in marrow cultured as an intact organ (marrow organ cultures), reduced mineralization was observed in mice given 149 Bq 241 Am/g mouse or more (skeletal dose rate of 25 mGy/day), in stromal marrow cells cultured from adherent cell layers and subsequently brought into a three-dimensional (3D) mineralizing condition (stromal 3D cultures), reduced "Sr uptake was observed from the lowest dose level tested (42 Bq 241 Am/g mouse, skeletal dose rate of 7 mGy/day) . Taking into account that only a fraction of the skeletal a-dose reached the marrow of the femoral diaphyses, marrow organ cultures and stromal 3D cultures exhibited high radiosensitivity to a-irradiation in vivo .However, after acute X-irradiation of marrow in vivo or in vitro prior to initiation of the marrow organ cultures, X-ray doses of 4 Gy or higher were needed to significantly impair the mineralization capacity of marrow organ cultures in vitro . Our data demonstrated that the osteogenic cells from the bone marrow are subjected to long-term damage after low doses of continuous a-irradiation in vivo .

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“…Several mouse or rat osteoblastic cell lines, representing various stages of osteoblast differentiation, are available, providing useful tools with which to study osteoblast-specific gene expression (29,31,44). Among them, the ROS cell lines derived from a rat osteosarcoma have been widely used (29).…”

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confidence: 99%

Two Distinct Osteoblast-Specific cis-Acting Elements Control Expression of a Mouse Osteocalcin Gene

Ducy

Karsenty

1995

Molecular and Cellular Biology

552

610

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Osteoblasts are cells of mesodermal origin that play a pivotal role during bone growth and mineralization. The mechanisms governing osteoblast-specific gene expression are still unknown. To understand these mechanisms, we analyzed the cis-acting elements of mouse osteocalcin gene 2 (mOG2), the best-characterized osteoblast-specific gene, by DNA transfection experiments in osteoblastic and nonosteoblastic cell lines and by DNA-binding assays. 5 deletion analysis of an mOG2 promoter-luciferase chimeric gene showed that a region located between ؊147 and ؊34 contained most if not all of the regulatory elements required for osteoblastspecific expression. Three different binding sites, called A, B, and C, for factors present in nuclear extracts of osteoblasts were identified in this short promoter by DNase I footprint assays. In gel retardation assays, the A element, located between bp ؊64 and ؊47, bound a factor present only in nuclear extracts of osteoblastic cell lines and nonmineralizing primary osteoblasts. The B element, located between bp ؊110 and ؊83, bound a ubiquitously expressed factor. The C element, located between bp ؊146 and ؊132, bound a factor present only in nuclear extracts of osteoblastic cell lines and nonmineralizing and mineralizing primary osteoblasts. When cloned upstream of a minimum osteocalcin promoter or a heterologous promoter, multimers of the A element strongly increased the activities of these promoters in osteoblastic cell lines at two different stages of differentiation but in no other cell line; we named this element osteocalcin-specific element 1 (OSE1). Multimers of the C element increased the activities of these promoters predominantly in a differentiated osteoblastic cell line; we named this element OSE2. This study demonstrates that two distinct cis-acting elements are responsible for osteoblast expression of mOG2 and provides for the first time a functional characterization of osteoblast-specific cis-acting elements. We speculate that these two elements may be important at several stages of osteoblast differentiation.

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Establishment of an osteogenic cell line derived from adult mouse bone marrow stroma by use of a recombinant retrovirus

Cited by 27 publications

References 38 publications

Development and Characterization of Conditionally Immortalized Osteoblast Precursor Cell Lines from Human Bone Marrow Stroma

Development and Characterization of Conditionally Immortalized Osteoblast Precursor Cell Lines from Human Bone Marrow Stroma

High Radiosensitivity of the Mineralization Capacity of Adult Murine Bone Marrowin Vitroto Continuous α-irradiation Compared to Acute X-irradiation

Two Distinct Osteoblast-Specific cis-Acting Elements Control Expression of a Mouse Osteocalcin Gene

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