Giemsa as a fluorescent stain for mineralized bone.

Bradbeer, Jeremy N.; Riminucci, Mara; Bianco, Paolo

doi:10.1177/42.5.7512588

Cited by 15 publications

(13 citation statements)

References 1 publication

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“…Bone of score 2 formed in the transplants of strains H and R was considerably more extensive than any bone, formed in vivo by several hESC lines, described thus far [3,5,19,20]. The new bone demonstrated numerous osteocytes embedded in lamellar, well-mineralized matrix, as can be judged by parallel organization of collagen bundles revealed under polarized light, and by intense green fluorescence revealed under ultraviolet light; such fluorescence was shown earlier to selectively distinguish mineralized bone in sections stained with eosincontaining dyes [28,29]. These results imply that long-term cultivation of hESC-derived strains in differentiating conditions employed in this study, including several weeks in postconfluent, multilayered cultures, followed by multiple passages and transplantation in HA=TCP vehicles, may, indeed, lead to the creation of clinically relevant bone tissue by nudging a subset of hESC descendants toward the osteogenic pathway.…”

Section: Figmentioning

confidence: 87%

In Vivo Bone Formation by Progeny of Human Embryonic Stem Cells

Kuznetsov

Cherman

Robey

2011

Stem Cells and Development

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The derivation of osteogenic cells from human embryonic stem cells (hESCs) or from induced pluripotent stem cells for bone regeneration would be a welcome alternative to the use of adult stem cells. In an attempt to promote hESC osteogenic differentiation, cells of the HSF-6 line were cultured in differentiating conditions in vitro for prolonged periods of time ranging from 7 to 14.5 weeks, followed by in vivo transplantation into immunocompromised mice in conjunction with hydroxyapatite=tricalcium phosphate ceramic powder. Twelve different medium compositions were tested, along with a number of other variables in culture parameters. In differentiating conditions, HSF-6-derived cells demonstrated an array of diverse phenotypes reminiscent of multiple tissues, but after a few passages, acquired a more uniform, fibroblast-like morphology. Eight to 16 weeks post-transplantation, a group of transplants revealed the formation of histologically proven bone of human origin, including broad areas of multiple intertwining trabeculae, which represents by far the most extensive in vivo bone formation by the hESC-derived cells described to date. Knockout-Dulbecco's modified Eagle's medium-based media with fetal bovine serum, dexamethasone, and ascorbate promoted more frequent bone formation, while media based on a-modified minimum essential medium promoted teratoma formation in 12-to 20-week-old transplants. Transcription levels of pluripotency-related (octamer binding protein 4, Nanog), osteogenesis-related (collagen type I, Runx2, alkaline phosphatase, and bone sialoprotein), and chondrogenesis-related (collagen types II and X, and aggrecan) genes were not predictive of either bone or teratoma formation. The most extensive bone was formed by the strains that, following 4 passages in monolayer conditions, were cultured for 23 to 25 extra days on the surface of hydroxyapatite=tricalcium phosphate particles, suggesting that coculturing of hESC-derived cells with osteoconductive material may increase their osteogenic potential. While none of the conditions tested in this study, and elsewhere, ensured consistent bone formation by hESC-derived cells, our results may elucidate further directions toward the construction of bone on the basis of hESCs or an individual's own induced pluripotent stem cells.

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Section: Figmentioning

confidence: 87%

In Vivo Bone Formation by Progeny of Human Embryonic Stem Cells

Kuznetsov

Cherman

Robey

2011

Stem Cells and Development

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“…The BSE images reveal the mineralized tissue regions and therefore lack the observation of the soft tissue layer formed around the implant. To observe the soft tissue and the bone lining cells, the histological sections were stained with 20% Giemsa stain solution [34]. The stained sample sections were observed using an optical light microscope (Leica DM RXA2).…”

Section: Histological Analysis and Environmental Scanning Electron MImentioning

confidence: 99%

Improving the osteointegration and bone–implant interface by incorporation of bioactive particles in sol–gel coatings of stainless steel implants

et al. 2010

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“…Haematoxylin and eosin (H&E)-stained sections were studied by confocal fluorescence microscopy, taking advantage of the known properties of eosin as a fluorochrome 22 in order to investigate the fine detail of histological changes observed in standard diagnostic material. Single focal plane confocal images or stacks of images spanning the entire section thickness were recorded and reconstructed in a Sarastro Laser Scanning Confocal System equipped with a 488 argon ion laser and complemented with the Molecular Dynamics Image Space software run on a Silicon Graphics workstation.…”

Section: Confocal Microscopymentioning

confidence: 99%