Dilution of human mesenchymal stem cells with dermal fibroblasts and the effects on in vitro and in vivo osteochondrogenesis

Lennon, Donald P.; Haynesworth, Stephen E.; Douglas, M.; Baber, Marilyn A.; Caplan, Arnold I.

doi:10.1002/1097-0177(2000)9999:9999<::aid-dvdy1037>3.0.co;2-7

Cited by 78 publications

(61 citation statements)

References 46 publications

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“…20,21 Human SFs, on the one hand, have been shown to be devoid of osteogenic potential. 22 However, in the recent study of Krebsbach et al 4 rat dermal fibroblasts revealed osteogenic potential when transduced with Ad5-BMP7 and subsequently subcutaneously implanted with gelatin sponges into immunocompromised mice. The new bone was formed within 1-3 weeks, and the newly formed bone was found to be a chimera composed of both human and mouse cells.…”

Section: Discussionmentioning

confidence: 98%

Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type

et al. 2003

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Section: Discussionmentioning

confidence: 98%

Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type

et al. 2003

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“…Also, freshly isolated ASCs can be contaminated with other cell types present in adipose tissue. The presence of these contaminating cells can affect ASC differentiation potential (29,30). For example, if fibroblasts make up a significant portion of the harvest population, then overall osteogenic potential could be significantly reduced.…”

Section: Discussionmentioning

confidence: 99%

Cellular mechanical properties reflect the differentiation potential of adipose-derived mesenchymal stem cells

González-Cruz

Fonseca²,

Darling³

2012

Proc. Natl. Acad. Sci. U.S.A.

194

153

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The mechanical properties of adipose-derived stem cell (ASC) clones correlate with their ability to produce tissue-specific metabolites, a finding that has dramatic implications for cell-based regenerative therapies. Autologous ASCs are an attractive cell source due to their immunogenicity and multipotent characteristics. However, for practical applications ASCs must first be purified from other cell types, a critical step which has proven difficult using surface-marker approaches. Alternative enrichment strategies identifying broad categories of tissue-specific cells are necessary for translational applications. One possibility developed in our lab uses single-cell mechanical properties as predictive biomarkers of ASC clonal differentiation capability. Elastic and viscoelastic properties of undifferentiated ASCs were tested via atomic force microscopy and correlated with lineage-specific metabolite production. Cell sorting simulations based on these "mechanical biomarkers" indicated they were predictive of differentiation capability and could be used to enrich for tissue-specific cells, which if implemented could dramatically improve the quality of regenerated tissues.cell mechanics | mesenchymal stem cell enrichment | viscoelasticity | single-cell characterization | AFM A dipose tissue contains a heterogeneous population of mesenchymal stem cells (MSCs) known as adipose-derived stem cells (ASCs) (1). ASCs are capable of differentiating into a variety of lineage-specific cell types, including adipocytes, osteoblasts, and chondrocytes (1-3). In comparison to MSCs derived from other tissues, ASCs are simple to isolate and available in large quantities (4, 5). Because of the cells' mesodermal origin, ASCs have been used for many soft tissue and orthopaedic applications (6-10). Unfortunately, ASC isolation is confounded by the lack of distinct and universally effective MSC biomarkers. Adipose tissue contains multiple cell types, including mature adipocytes, fibroblasts, smooth muscle cells, and endothelial cells (11), which can contaminate the stromal fraction collected during ASC isolation. While conventional methods such as flow cytometry can isolate stem cells using surface antigen expression (1, 2, 12), resulting cell yields are often less than 1% (13,14). Furthermore, the surface antigens present on ASCs can also be found on other cell types in adipose tissue, complicating the isolation of pure mesenchymal stem cell populations (15-17). Collectively, these limitations suggest a need for alternative biomarkers that allow for ASC enrichment based on lineage potential.Recently, single-cell mechanical properties were found to be akin to gene and protein expressions, capable of distinguishing differences in cellular subpopulations, disease state, and tissue source (18)(19)(20)(21)(22). Cells display varying levels of resistance to deformation (elasticity) and flow (viscosity) in response to an applied force. This behavior depends on the composition and organization of subcellular structures, particularly the cytosk...

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“…The pore structure ensures intimate cell-cell contact, which should enhance chondrogenic differentiation. 32,33 Static seeding (i.e., the cells are layered on to the scaffold) has historically been the most commonly used seeding method. 14,34-37 Drawbacks of this method are that, as in our study, it usually results in a relatively low initial cell density.…”

Section: Discussionmentioning

confidence: 99%

A Rapid Seeding Technique for the Assembly of Large Cell/Scaffold Composite Constructs

Solchaga¹,

Tognana²,

Penick³

et al. 2006

Tissue Engineering

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These studies address critical technical issues involved in creating human mesenchymal stem cell (hMSC)/scaffold implants for cartilage repair. These issues include obtaining a high cell density and uniform spatial cell distribution within the scaffold, factors that are critical in the initiation and homogeneity of chondrogenic differentiation. For any given scaffold, the initial seeding influences cell density, retention, and spatial distribution within the scaffold, which eventually will affect the function of the construct. Here, we discuss the development of a vacuum-aided seeding technique for HYAFF ® -11 sponges which we compared to passive infiltration. Our results show that, under the conditions tested, hMSCs were quantitatively and homogeneously loaded into the scaffolds with 90+% retention rates after 24 h in perfusion culture with no negative effect on cell viability or chondrogenic potential. The retention rates of the vacuum-seeded constructs were at least 2 times greater than those of passively seeded constructs at 72 h. Histomorphometric analysis revealed that the core of the vacuum-seeded constructs contained 240% more cells than the core of passively infiltrated scaffolds. The vacuum seeding technique is safe, rapid, reproducible, and results in controlled quantitative cell loading, high retention, and uniform distribution.

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Dilution of human mesenchymal stem cells with dermal fibroblasts and the effects on in vitro and in vivo osteochondrogenesis

Cited by 78 publications

References 46 publications

Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type

Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type

Cellular mechanical properties reflect the differentiation potential of adipose-derived mesenchymal stem cells

A Rapid Seeding Technique for the Assembly of Large Cell/Scaffold Composite Constructs

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