Characterization of endosteal osteoblastic cells isolated from mouse caudal vertebrae

Lomri, Abderrahim; Marie, P.; Tran, Philippe; Hott, M.

doi:10.1016/8756-3282(88)90006-3

Cited by 61 publications

(32 citation statements)

References 32 publications

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“…This was carried out according to previously reported methods (28). The thermosensitivity of the enzyme was checked according to Goldstein et al (12).…”

Section: Biochemical Measurements Of Alkaline Phosphatase Activitymentioning

confidence: 99%

“…Cells and nodule sections were exposed to naphthol ASBI phosphate (30 min, 370C, pH 8.6) and naphthol ASTR (Sigma Chemical Co.) (1 h, 37°C, pH 6), used as substrates for alkaline and acid phosphatase activities, respectively (28). The cells were observed with or without counterstaining with hematoxylin.…”

Section: Phosphatase Activitiesmentioning

confidence: 99%

“…5-p,m-thick sections were stained with toluidine blue, Von Kossa, or Goldner Trichrome for the detection of collagen and minerals (2,28).…”

Section: Histological Detection Of Collagen and Mineralsmentioning

confidence: 99%

“…(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.…”

mentioning

confidence: 99%

“…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.…”

mentioning

confidence: 99%

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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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“…This was carried out according to previously reported methods (28). The thermosensitivity of the enzyme was checked according to Goldstein et al (12).…”

Section: Biochemical Measurements Of Alkaline Phosphatase Activitymentioning

confidence: 99%

Section: Phosphatase Activitiesmentioning

confidence: 99%

“…5-p,m-thick sections were stained with toluidine blue, Von Kossa, or Goldner Trichrome for the detection of collagen and minerals (2,28).…”

Section: Histological Detection Of Collagen and Mineralsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

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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Proliferation and differentiation of human trabecular osteoblastic cells on hydroxyapatite

Hott

Noël

Bernache-Assolant

et al. 1997

J. Biomed. Mater. Res.

118

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In order to evaluate whether human osteoblastic cells differentiate normally on hydroxyapatite, we have compared the adhesion, proliferation, and differentiation of human trabecular (HT) osteoblastic cells on synthetic-dense hydroxyapatite and on standard plastic culture. We show here that initial HT cell attachment was 4-fold lower on hydroxyapatite than on plastic after 4 h of culture, and that normal cell attachment on hydroxyapatite was restored after 18 h of culture. HT cell proliferation was similar on the two substrates at 2-8 days of culture, but was lower on hydroxyapatite compared to plastic after 15 and 28 days of culture, as evaluated by DNA synthesis or cell number. HT cells cultured on both substrates produced an abundant extracellular matrix which immunostained for Type I collagen. The levels of carboxyterminal propeptide of Type I procollagen (P1CP) in the medium were lower in HT cell cultures on hydroxyapatite than on plastic. In addition, (3H)-proline incorporation into matrix proteins and the mean thickness of matrix layers were 52% and 26% lower, respectively, on hydroxyapatite compared to plastic after 4 weeks of culture, indicating that the total collagenous matrix synthesized by HT cells was lower on hydroxyapatite. However, (3H)-proline and calcium uptake expressed per cell was higher on hydroxyapatite than on plastic. The results show that human osteoblastic cells attach, proliferate, and differentiate on dense hydroxyapatite with a sequence similar to that of plastic. However, the growth of human osteoblastic cells is lower on hydroxyapatite in long-term culture, which results in a reduced amount of extracellular matrix, although matrix production per cell may be increased.

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Anin vitro system for studying osteointegration of dental implants utilizing cells grown on dense hydroxyapatite disks

Layman

Ardoin

1998

J. Biomed. Mater. Res.

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A proposed in vitro system is described where chick osteoblasts are cultured on the flat surfaces of dense, nonporous HA disks to facilitate the study of bone formation at the cell-HA interface. During early bone formation cell-coated HA disks were retrieved, fixed with buffered 2% glutaraldehyde, and embedded in epon/araldite. The underlying HA disks were demineralized in diluted acid, and the intact cell-HA interfaces were re-embedded and thin sectioned for routine transmission electron microscopy. Morphologic studies indicated that osteoblasts proliferated and formed nodules of cells on the surfaces of HA disks. With increasing time in culture, they deposited orthogonally packed collagen fibrils between the cell layers that were enveloped by electron-dense mineralized globules. Eventually, small spicules of mineralized HA formed along collagen fibrils. An electron-dense layer about 50 nm thick was observed on the surface of the HA disks. Biochemical studies indicated that cell proliferation, as judged by 3H-thymidine uptake, increased rapidly during the first 3 days, reached a maximum around 6 days, and then declined by 12 days in culture. AP activity and collagen synthesis, as determined by 3H-hydroxyproline formation, increased as cellular proliferation declined. Mineralization, as judged by 45Ca uptake and spicule formation, occurred, as expected, following the increase in AP activity and deposition of densely packed collagen fibrils. Thus, all morphological and biochemical parameters studied indicate that the proposed in vitro system is reproducible and can facilitate the study of the osteointegration of HA-coated implants.

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Characterization of endosteal osteoblastic cells isolated from mouse caudal vertebrae

Cited by 61 publications

References 32 publications

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

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

Proliferation and differentiation of human trabecular osteoblastic cells on hydroxyapatite

Anin vitro system for studying osteointegration of dental implants utilizing cells grown on dense hydroxyapatite disks

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