Bone formation in bone marrow organ cultures

Luriya, E. A.; Owen, Maureen; Fridenshtein, A. Ya.; Kuznetsov, Sergei A.; Genkina, E. N.; Gosteva, V. V.

doi:10.1007/bf00834432

Cited by 11 publications

(12 citation statements)

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“…It is unclear why this area does not induce endochondraltype ossification, but it is worthy of note that chondrocytes had degenerated prior to vessel invasion accompanying osteogenic cells. In the present organ culture, it is likely that fl-glycerophosphate acted as a enhancer of ALPase activity (Luria et al 1987;Maniatopoulos et al 1988;Benayahu et al 1989) in transforming cells forming organized extracellular matrix, and thereby might facilitate matrix calcification. These effects of fl-glycerophosphate on calcification were observed similarly in the extracellular matrix containing not only type I collagen but also type II collagen, whereas since the transformation in chondrocytes occurred independently of matrix calcification, there was no evidence that the calcification-eliciting effect with fl-glycerophosphate acted as a direct trigger for the cellular transformation.…”

Section: Discussionmentioning

confidence: 99%

Meckel's cartilage chondrocytes in organ culture synthesize bone-type proteins accompanying osteocytic phenotype expression

et al. 1996

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We examined whether Meckel's cartilage of embryonic mice, 17 days in utero, undergo the cellular transformation into the osteocyte-like phenotype under organ culture conditions. Explants were grown by our original pithole method modified Trowell-type cultures for up to 4 weeks at 37 degrees C under 5% CO2 in air. Specimens were examined using histological procedures including immunostaining and electron microscopy. In addition, the effects of beta-glycerophosphate on matrix calcification were also examined in cultures with or without beta-glycerophosphate. Addition of beta-glycerophosphate induced calcification at a higher level, but calcium mineral deposition occurred regardless of the addition of beta-glycerophosphate to the culture medium. Light and electron microscopic analyses showed that freshly isolated chondrocytes prior to cell culture had typical hypertrophic morphology, but shortly after commencement of culture, they showed morphological modifications. The cells showing chondrocytic phenotypes became basophilic elliptical cells, and eventually transformed into flattened osteocyte-like cells. Bone-like features for cellular elements were characterized by spindle-shaped cells with elongated processes accompanying bone-specific thick-banded collagen fibrils. Immunostaining showed that at 2 weeks in culture, type I and type II collagens coexisted in the matrix, but subsequently type II collagen synthesis ceased and was replaced by type I collagen synthesis. Immunofluorescent labeling for osteocalcin was noted first in the peripheral cells by 1 week, but at 3 weeks this reaction spread to the central zone in explants. Alkaline phosphatase activity (ALPase) was expressed on the cells in the central zone prior to calcium mineral deposition as shown by von Kossa's reaction at 3 weeks in culture. These results showed that Meckel's cartilage chondrocytes in organ culture synthesize bone-type proteins accompanying osteocytic phenotype expression.

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Section: Discussionmentioning

confidence: 99%

Meckel's cartilage chondrocytes in organ culture synthesize bone-type proteins accompanying osteocytic phenotype expression

et al. 1996

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“…Bone marrow stromal cells have multilineage differentiation potential and can differentiate into cells of mesenchymal origin with an osteogenic phenotype. Accordingly, they have been frequently used for bone reconstruction (Luria et al 1987;Haynesworth et al 1992;Matsubara et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Effects of FGF2 and FGF9 on osteogenic differentiation of bone marrow-derived progenitors

Kizhner

Ben-David

Rom

et al. 2011

In Vitro Cell.Dev.Biol.-Animal

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Bone repair is a major concern in reconstructive surgery. Transplants containing osteogenically committed mesenchymal stem cells (MSCs) provide an alternative source to the currently used autologous bone transplants which have limited supply and require additional surgery to the patient. A major drawback, however is the lack of a critical mass of cells needed for successful transplantation. The purpose of the present study was to test the effects of FGF2 and FGF9 on expansion and differentiation of MSCs in order to establish an optimal culture protocol resulting in sufficient committed osteogenic cells required for successful in vivo transplantation. Bone marrow-derived MSCs cultured in αMEM medium supplemented with osteogenic supplements for up to three passages (control medium), were additionally treated with FGF2 and FGF9 in various combinations. Cultures were evaluated for viability, calcium deposition and in vivo osteogenic capacity by testing subcutaneous transplants in nude mice. FGF2 had a positive effect on the proliferative capacity of cultured MSCs compared to FGF9 and control medium treated cultures. Cultures treated with FGF2 followed by FGF9 showed an increased amount of extracted Alizarin red indicating greater osteogenic differentiation. Moreover, the osteogenic capacity of cultured cells transplanted in immunodeficient mice revealed that cells that were subjected to treatment with FGF2 in the first two passages and subsequently to FGF9 in the last passage only, were more successful in forming new bone. It is concluded that the protocol using FGF2 prior to FGF9 is beneficial to cell expansion and commitment, resulting in higher in vivo bone formation for successful bone tissue engineering.

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“…More recently, in vitro systems have been developed in which mineralized bone develops from whole bone marrow (Luria et al 1987 or from a stomal adherent cell population (Maniatopoulos 1988, Long et al . 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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Bone formation in bone marrow organ cultures

Cited by 11 publications

References 1 publication

Meckel's cartilage chondrocytes in organ culture synthesize bone-type proteins accompanying osteocytic phenotype expression

Meckel's cartilage chondrocytes in organ culture synthesize bone-type proteins accompanying osteocytic phenotype expression

Effects of FGF2 and FGF9 on osteogenic differentiation of bone marrow-derived progenitors

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

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