Regionally-derived cell populations and skeletal stem cells from human foetal femora exhibit specific osteochondral and multi-lineage differentiation capacity in vitro and ex vivo

Gothard, David; Cheung, Kelvin; Kanczler, Janos M.; Wilson, David I.; Oreffo, Richard O.C.

doi:10.1186/s13287-015-0247-2

Cited by 12 publications

(17 citation statements)

References 65 publications

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“…These results are in agreement with previous reports. In [ 10 ], the percentage of Stro-1 + cells in adult human BM cells declined to 8.5% after six weeks in culture, and in [ 22 ], Stro-1 + cells were completely absent at passage 7, after 21–35 DIV, in human fetal femur-derived BM cell cultures. Stewart et al .…”

Section: Resultsmentioning

confidence: 99%

“…ALP activity was quantified against a standard curve (p-nitrophenol) by incubation of the cell lysate with a phosphatase substrate solution (1.33 mg ml −1 p -nitrophenyl phosphate in 0.5 M alkaline buffer solution) at 37°C in the dark, under gentle agitation. The reaction was terminated by addition of 1 M NaOH in dH 2 O after 40 min and absorbance was measured at 410 nm using an ELx800™ absorbance reader (BioTek) [ 22 , 45 ].…”

Section: Methodsmentioning

confidence: 99%

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Size and dielectric properties of skeletal stem cells change critically after enrichment and expansion from human bone marrow: consequences for microfluidic cell sorting

Martín

Andrés

Spencer

et al. 2017

J. R. Soc. Interface.

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The capacity of bone and cartilage to regenerate can be attributed to skeletal stem cells (SSCs) that reside within the bone marrow (BM). Given SSCs are rare and lack specific surface markers, antibody-based sorting has failed to deliver the cell purity required for clinical translation. Microfluidics offers new methods of isolating cells based on biophysical features including, but not limited to, size, electrical properties and stiffness. Here we report the characterization of the dielectric properties of unexpanded SSCs using single-cell microfluidic impedance cytometry (MIC). Unexpanded SSCs had a mean size of 9.0 µm; larger than the majority of BM cells. During expansion, often used to purify and increase the number of SSCs, cell size and membrane capacitance increased significantly, highlighting the importance of characterizing unaltered SSCs. In addition, MIC was used to track the osteogenic differentiation of SSCs and showed an increased membrane capacitance with differentiation. The electrical properties of primary SSCs were indistinct from other BM cells precluding its use as an isolation method. However, the current studies indicate that cell size in combination with another biophysical parameter, such as stiffness, could be used to design label-free devices for sorting SSCs with significant clinical impact.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Size and dielectric properties of skeletal stem cells change critically after enrichment and expansion from human bone marrow: consequences for microfluidic cell sorting

Martín

Andrés

Spencer

et al. 2017

J. R. Soc. Interface.

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“…These alloys also promote the osteogenic differentiation of hAMSCs (Marini et al, ). In addition, a recent study by Gothard et al () has shown that regionally derived cell populations and skeletal stem cells from human fetal femora exhibit specific osteochondral and multi‐lineage differentiation capacity both ex vivo and in culture.…”

Section: Introductionmentioning

confidence: 99%

Stem cells applications in bone and tooth repair and regeneration: New insights, tools, and hopes

Meguid

et al. 2017

Journal Cellular Physiology

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The exploration of stem and progenitor cells holds promise for advancing our understanding of the biology of tissue repair and regeneration mechanisms after injury. This will also help in the future use of stem cell therapy for the development of regenerative medicine approaches for the treatment of different tissue-species defects or disorders such as bone, cartilages, and tooth defects or disorders. Bone is a specialized connective tissue, with mineralized extracellular components that provide bones with both strength and rigidity, and thus enable bones to function in body mechanical supports and necessary locomotion process. New insights have been added to the use of different types of stem cells in bone and tooth defects over the last few years. In this concise review, we briefly describe bone structure as well as summarize recent research progress and accumulated information regarding the osteogenic differentiation of stem cells, as well as stem cell contributions to bone repair/regeneration, bone defects or disorders, and both restoration and regeneration of bones and cartilages. We also discuss advances in the osteogenic differentiation and bone regeneration of dental and periodontal stem cells as well as in stem cell contributions to dentine regeneration and tooth engineering.

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“…Further, a direct comparison of fetal chondroprogenitors from different origins, such as femoral head, spine and ulnar epiphysis (used in this study), could determine the most stable cell source for cartilage regeneration. Reports by others suggest that cells isolated from fetal femoral head progress towards hypertrophic cartilage in vitro (Quintin et al , ; Gothard et al , ).…”

Section: Discussionmentioning

confidence: 99%

Human chondroprogenitors in alginate-collagen hybrid scaffolds produce stable cartilagein vivo

Studer

Cavalli

Formica

et al. 2016

J Tissue Eng Regen Med

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The goal of this study was to evaluate human epiphyseal chondroprogenitor cells (ECPs) as a potential new cell source for cartilage regeneration. ECPs were compared to human bone marrow stromal cells (MSCs) and human adult articular chondrocytes (ACs) for their chondrogenic potential and phenotypic stability in vitro and in vivo. The cells were seeded in Optimaix-3D scaffolds at 5 × 10 cells/mm and gene expression, matrix production and mechanical properties were analysed up to 6 weeks. In vitro, ECPs synthesized consistently high collagen 2 and low collagen 10. AC-seeded constructs exhibited high donor variability in GAG/DNA values as well as in collagen 2 staining, but showed low collagen 10 production. MSCs, on the other hand, expressed high levels of collagen 2 but also of collagens 1 and 10, and were therefore not considered further. In vivo, there was considerable loss of matrix proteins in ECPs compared to in vitro cultured samples. To overcome this, a second implantation study investigated the effect of mixing cells with alginate prior to seeding in the scaffold. ECPs in alginate maintained their cartilage matrix and resisted mineralization and vessel infiltration better 6 weeks after subcutaneous implantation, whereas ACs lost their chondrogenic matrix completely. This study shows the great potential of ECPs as an off-the-shelf, highly chondrogenic cell type that produces stable cartilage in vivo. Copyright © 2016 John Wiley & Sons, Ltd.

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Regionally-derived cell populations and skeletal stem cells from human foetal femora exhibit specific osteochondral and multi-lineage differentiation capacity in vitro and ex vivo

Cited by 12 publications

References 65 publications

Size and dielectric properties of skeletal stem cells change critically after enrichment and expansion from human bone marrow: consequences for microfluidic cell sorting

Size and dielectric properties of skeletal stem cells change critically after enrichment and expansion from human bone marrow: consequences for microfluidic cell sorting

Stem cells applications in bone and tooth repair and regeneration: New insights, tools, and hopes

Human chondroprogenitors in alginate-collagen hybrid scaffolds produce stable cartilagein vivo

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