Bone formation by osteoblast-like cells in a three-dimensional cell culture

Casser-Bette, M; Murray, Andrew B.; Closs, Ellen I.; Erfle, Volker; Schmidt, Jörg

doi:10.1007/bf02555824

Cited by 119 publications

(63 citation statements)

References 29 publications

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“…Interestingly, while osteoblasts migrated into the gels and formed a 3D cell network, fibroblasts seeded on top of the gels migrated through them and formed a monolayer on the plastic underneath. 7 In our experiments the cells were mixed with the collagen solution before it set as a gel, and hence the cells settled directly into the 3D environment. An inherent methodological difficulty in the study of cells in 2D vs 3D cultures is the fact that the cell densities in the two growth environments are not directly comparable.…”

Section: Enhanced Osteoblastogenesis In 3d Gelsmentioning

confidence: 99%

“…4 Although a number of studies have investigated osteoblasts cultured in 3D collagen gels, they mostly used microscopy for visual analysis of the osteoblast phenotype and alkaline phosphatase activity as a measure of osteoblast function. 1,3,[7][8][9] The aim of the current study was to directly compare osteoblasts growing in parallel in 3D collagen gels and in 2D on plastic and to comprehensively characterize the effects of these growth environments on the expression of osteoblast marker genes, on cell differentiation and on responses to factors that affect the rate of cell proliferation.…”

Section: Introductionmentioning

confidence: 99%

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Enhanced osteoblastogenesis in three-dimensional collagen gels

et al. 2014

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Growth and differentiation of osteoblasts are often studied in cell cultures. In vivo, however, osteoblasts are embedded within a complex three-dimensional (3D) microenvironment, which bears little relation to standard culture flasks. Our study characterizes osteoblast-like cells cultured in 3D collagen gels and compares them with cells in two-dimensional (2D) cultures. Primary rat osteoblasts and MC3T3-E1 cells were seeded within type I collagen gels, and differentiation was determined by mineral staining and gene expression analysis. Cells growing in 3D gels showed positive mineral staining and induction of osteoblast marker genes earlier than cells growing in 2D. A number of genes, including osteocalcin, bone sialoprotein, alkaline phosphatase and dentin matrix protein 1, were already highly upregulated in 3D cultures 24 h after seeding. The early expression of osteoblast genes was dependent on the 3D structure and was not induced in cells growing on collagen-coated dishes in 2D. Comparison of thymidine incorporation between cells in 3D and 2D cultures treated with agents that induce proliferation-transforming growth factor b, platelet-derived growth factor and lactoferrin-showed a much greater response in 3D gels. Cells in 3D cultures were also much more sensitive to inhibition of proliferation by the protein kinase inhibitor imatinib mesylate. The 3D collagen gels better represent the physiological bone environment and offer a number of technical advantages for the study of osteoblasts in vitro. These studies have additional practical implications as 3D collagen gels are considered as a scaffold material in regenerative medicine for the repair of bone defects.

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Section: Enhanced Osteoblastogenesis In 3d Gelsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Enhanced osteoblastogenesis in three-dimensional collagen gels

et al. 2014

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“…These were based on the utilization of supports, as gels and sponges of collagen type I, PLGA foam, DegraPol, agarose, Matrigel, or methylcellulose. 7,10,15,16,18,20,22,25,26,30,32,34,37,39 Normal human osteogenic cells, osteoblasts from prepubertal-age donors, or bone marrow stromal fibroblasts adhering to scaffolding of HAP/ TC, 16,20,25,26,30,34 collagraft, 26 collagen sponges, 15,39 or bone chips 5 were utilized to produce specifically 3D structures designed for direct implantation in vivo.…”

Section: Introductionmentioning

confidence: 99%

Three-dimensional cultures of normal human osteoblasts: proliferation and differentiation potential in vitro and upon ectopic implantation in nude mice

Ferrera

Poggi

Biassoni

et al. 2002

Bone

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“…The majority of these studies have utilized an osteoblastic cell line in culture media that typically contain ionic calcium levels between 1.8 mM [Minimum Essential Medium (MEM), aMEM, and Dulbecco's Modified Eagle Medium (DMEM)] and 2.6 mM [Biggers, Gwatkin and Judah b (BGJ/b) Medium, Fitton Jackson Modification], ionic phosphate levels of 0.9 mM (DMEM) to 1.6 mM (BGJ/b), and 10 mM b-glycerophosphate [1][2][3][4][5][6][7][8][9][10][11][12][13][14]. The role of the bglycerophosphate is generally accepted to be as a source of phosphate for mineral growth, but it may affect mineral deposition in other ways as well [15].…”

mentioning

confidence: 99%

Mineralization of Decalcified Bone Occurs Under Cell Culture Conditions and Requires Bovine Serum But Not Cells

Hamlin

Price

2004

Calcif Tissue Int

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Abstract. The purpose of this study was to develop an in vitro model system for bone matrix mineralization in the absence of cells. For this model, we utilized EDTAdecalcified new-born rat tibias with the cartilaginous ends intact, allowing us to visually determine the specificity of mineralization within the bone. Our results show that supplementation of DMEM culture medium with 10mM b-glycerophosphate and 15% fetal bovine serum (FBS) results in non-physiological mineral percipitation in the tibia because of the generation of supraphysiological (5mM) levels of inorganic phosphate in the medium. The same medium supplemented only with inorganic phosphate to a final concentration of 2mM failed to mineralize a decalcified tibia matrix. However, additional supplementation of this medium with as little as 5% FBS resulted in mineralization of those regions of the type I collagen where mineral was found prior to decalcification, with no evidence for mineralization in the cartilage at the bone ends or in the periosteum. Analysis of the mineral by Fourier-transform infrared spectroscopy and powder X-ray diffraction shows that tibias that have been decalcified and then remineralized contain an apatitic mineral that is strikingly similar to the mineral in normal bone. Tendon, a type I collagen matrix not normally mineralized in vivo , also mineralizes when incubated in DMEM containing 2mM Pi and as little as 1.5% FBS, but not when incubated in DMEM without serum. These data indicate that serum contains a nucleator of type I collagen matrix mineralization, and that mineralization of type I collagen under cell culture conditions requires serum but not living cells.

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Bone formation by osteoblast-like cells in a three-dimensional cell culture

Cited by 119 publications

References 29 publications

Enhanced osteoblastogenesis in three-dimensional collagen gels

Enhanced osteoblastogenesis in three-dimensional collagen gels

Three-dimensional cultures of normal human osteoblasts: proliferation and differentiation potential in vitro and upon ectopic implantation in nude mice

Mineralization of Decalcified Bone Occurs Under Cell Culture Conditions and Requires Bovine Serum But Not Cells

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