Differentiation of Osteoblasts and<i>in Vitro</i>Bone Formation from Murine Embryonic Stem Cells

Buttery, Lee D.K.; Bourne, S.; Xynos, J. D.; Wood, H.; Hughes, FJ; Hughes, S. P. F.; Episkopou, Vasso; Polak, Julia M.

doi:10.1089/107632700300003323

Cited by 368 publications

(263 citation statements)

References 46 publications

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“…Dexamethasone augmented the nodule number dramatically, when added to ascorbic acid and h-glycerophosphate. Comparable results were obtained with murine embryonic stem cells [70]. In primary rat calvaria cell cultures, additional nodule formation was observed, and the self-renewal capacity of the bone-nodule forming cells was increased [71].…”

Section: Discussionmentioning

confidence: 58%

Fetal bone cells for tissue engineering

Montjovent

Burri

Mark

et al. 2004

Bone

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We envision the use of human fetal bone cells for engineered regeneration of adult skeletal tissue. A description of their cellular function is then necessary. To our knowledge, there is no description of human primary fetal bone cells treated with differentiation factors. The characterization of fetal bone cells is particularly important as the pattern of secreted proteins from osteoblasts has been shown to change during aging. In the first part of this work, human primary fetal bone cells were compared to adult bone cells and mesenchymal stem cells for their ability to proliferate and to differentiate into osteoblasts in vitro. Cell proliferation, gene expression of bone markers, alkaline phosphatase (ALP) activity, and mineralization were analyzed during a time-course study. In the second part of this paper, bone fetal cells behavior exposed to osteogenic factors is further detailed. The doubling time of fetal bone cells was comparable to mesenchymal stem cells but significantly shorter than for adult bone cells. Gene expression of cbfa-1, ALP, a1 chain of type I collagen, and osteocalcin were upregulated in fetal bone cells after 12 days of treatment, with higher inductions than for adult and mesenchymal stem cells. The increase of ALP enzymatic activity was stronger for fetal than for adult bone cells reaching a maximum at day 10, but lower than for mesenchymal stem cells. Importantly, the mineralization process of bone fetal cells started earlier than adult bone and mesenchymal stem cells. Proliferation of fetal and adult bone cells was increased by dexamethasone, whereas 1a,25-dihydroxyvitamin D 3 did not show any proliferative effect. Mineralization studies clearly demonstrated the presence of calcium deposits in the extracellular matrix of fetal bone cells. Nodule formation and calcification were strongly increased by the differentiation treatment, especially by dexamethasone. This study shows for the first time that human primary fetal bone cells could be of great interest for bone research, due to their fast growth rate and their ability to differentiate into mature osteoblasts. They represent an interesting and promising potential for therapeutic use in bone tissue engineering.

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Section: Discussionmentioning

confidence: 58%

Fetal bone cells for tissue engineering

Montjovent

Burri

Mark

et al. 2004

Bone

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“…However, up to 80% of the whole EB became necrotic by day 9 in all samples. This demonstrates a serious problem with extended culture C5 days which is the current conventional culture period (Buttery et al 2001;Yirme et al 2008). A variety of alternative avenues could be explored to resolve this problem including vascularization of the EB structure (Feraud et al 2001;Goodwin 2007).…”

Section: Discussionmentioning

confidence: 99%

Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

et al. 2009

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Cell-cell interaction is an integral part of embryoid body (EB) formation controlling 3D aggregation. Manipulation of embryonic stem (ES) cell interactions could provide control over EB formation. Studies have shown a direct relationship between EB formation and ES cell differentiation. We have previously described a cell surface modification and cross-linking method for influencing cell-cell interaction and formation of multicellular constructs. Here we show further characterisation of this engineered aggregation. We demonstrate that engineering accelerates ES cell aggregation, forming larger, denser and more stable EBs than control samples, with no significant decrease in constituent ES cell viability. However, extended culture C5 days reveals significant core necrosis creating a layered EB structure. Accelerated aggregation through engineering circumvents this problem as EB formation time is reduced. We conclude that the proposed engineering method influences initial ES cell-ES cell interactions and EB formation. This methodology could be employed to further our understanding of intrinsic EB properties and their effect on ES cell differentiation.

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“…Some spontaneous osteogenic differentiation of ESCs was observed in basic medium [127]. By using similar approaches as described above for chondrogenic differentiation, the osteogenic fraction could be increased.…”

Section: Growth Factor-induced Osteogenic Differentiation Of Escsmentioning

confidence: 90%

“…Growth factors and cytokines widely used to promote osteogenic differentiation of primary osteoblasts, pre-osteoblasts and adult stem cells [128] are AA, ß-glycerophosphate (BGP) and Dex. Supplementing ESC medium with AA and BGP [127] or the trio of supplements [129][130][131][132] significantly increased the amount of bone nodules and the expression of osteogenic markers in mouse and human ESC cultures. Osteogenic differentiation was also observed when intact EBs, rather than dissociated EB cells, were cultured in the presence of osteogenic supplements [133,134].…”

Section: Growth Factor-induced Osteogenic Differentiation Of Escsmentioning

confidence: 98%