Cartilage‐like gene expression in differentiated human stem cell spheroids: A comparison of bone marrow–derived and adipose tissue–derived stromal cells

Winter, Anja; Breit, Stephen; Parsch, Dominik; Benz, Karin; Steck, Eric; Hauner, Hans; Weber, R.; Ewerbeck, Volker; Richter, Wiltrud

doi:10.1002/art.10767

Cited by 417 publications

(337 citation statements)

References 64 publications

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“…Stem cells were proposed as an attractive alternate cell source because of their promising growth and differen- tiation potential and their ability to form cartilage-like tissue upon chondrogenic induction in vitro (10,36). Since functional suitability and phenotypic stability of the transplanted cells are crucial traits for the clinical application of MSCs in cartilage repair, the recapitulation of natural differentiation programs during chondrogenesis of MSCs and the adoption of a stable chondrocyte phenotype are desired.…”

Section: Discussionmentioning

confidence: 99%

Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice

Pelttari¹,

Winter²,

Steck³

et al. 2006

Arthritis & Rheumatism

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726

669

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Objective. Functional suitability and phenotypic stability of ectopic transplants are crucial factors in the clinical application of mesenchymal stem cells (MSCs) for articular cartilage repair, and might require a stringent control of chondrogenic differentiation. This study evaluated whether human bone marrow-derived MSCs adopt natural differentiation stages during induction of chondrogenesis in vitro, and whether they can form ectopic stable cartilage that is resistant to vascular invasion and calcification in vivo.Methods. During in vitro chondrogenesis of MSCs, the expression of 44 cartilage-, stem cell-, and bone-related genes and the deposition of aggrecan and types II and X collagen were determined. Similarly treated, expanded articular chondrocytes served as controls. MSC pellets were allowed to differentiate in chondrogenic medium for 3-7 weeks, after which the chondrocytes were implanted subcutaneously into SCID mice; after 4 weeks in vivo, samples were evaluated by histology.Results. The 3-stage chondrogenic differentiation cascade initiated in MSCs was primarily characterized by sequential up-regulation of common cartilage genes. Premature induction of hypertrophy-related molecules (type X collagen and matrix metalloproteinase 13) occurred before production of type II collagen and was followed by up-regulation of alkaline phosphatase activity. In contrast, hypertrophy-associated genes were not induced in chondrocyte controls. Whereas control chondrocyte pellets resisted calcification and vascular invasion in vivo, most MSC pellets mineralized, in spite of persisting proteoglycan and type II collagen content.Conclusion. An unnatural pathway of differentiation to chondrocyte-like cells was induced in MSCs by common in vitro protocols. MSC pellets transplanted to ectopic sites in SCID mice underwent alterations related to endochondral ossification rather than adopting a stable chondrogenic phenotype. Further studies are needed to evaluate whether a more stringent control of MSC differentiation to chondrocytes can be achieved during cartilage repair in a natural joint environment.

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Section: Discussionmentioning

confidence: 99%

Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice

Pelttari¹,

Winter²,

Steck³

et al. 2006

Arthritis & Rheumatism

Self Cite

726

669

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“…The most commonly used chondrogenic cocktail for ASCs (TGF-β1 and dexamethasone) was used for this study (Afizah et al 2007;Mehlhorn et al 2006;Winter et al 2003). However, genes not associated with chondrogenesis, per se, were upregulated.…”

Section: Discussionmentioning

confidence: 99%

Monolayer cell expansion conditions affect the chondrogenic potential of adipose‐derived stem cells

Estes

Diekman

Guilak

2008

Biotech & Bioengineering

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Adipose-derived stem cells (ASCs) are an abundant, readily available population of multipotent progenitor cells that reside in adipose tissue. Isolated ASCs are typically expanded in monolayer on standard tissue culture plastic with a basal medium containing 10% fetal bovine serum. However, recent data suggests that altering the monolayer expansion conditions by using suspension culture plastic, adding growth factors to the medium, or adjusting the seeding density may affect the self-renewal rate, multipotency, and lineage-specific differentiation potential of the ASCs. We hypothesized that variation in any of these expansion conditions would influence the chondrogenic potential of ASCs. ASCs were isolated from human liposuction waste tissue and expanded through two passages with different tissue culture plastic, feed medium, and cell seeding densities. Once expanded, the cells were cast in an agarose gel and subjected to identical chondrogenic culture conditions for 7 days, at which point cell viability, radiolabel incorporation, and gene expression were measured. High rates of matrix synthesis upon chondrogenic induction were mostly associated with smaller cells, as indicated by cell width and area on tissue culture plastic, and it appears that expansion in a growth factor supplemented medium is important in maintaining this morphology. All end-point measures were highly dependent on the specific monolayer culture conditions. These results support the hypothesis that monolayer culture conditions may "prime" the cells or predispose them towards a specific phenotype and thus underscore the importance of early culture conditions in determining the growth and differentiation potential of ASCs.

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“…For culturing MSC were replated at a density of 4-6 x10 3 cells/cm 2 . Cell surface marker expression profiles and the multipotency were standardly determined for MSC populations, as described previously (Winter et al, 2003;Dickhut et al, 2009). Depending on the experimental setting, cells from passage 1, 2, 3 or 5 were used.…”

Section: Cell Isolation and Cultivationmentioning

confidence: 99%

Prediction of in vivo bone forming potency of bone marrow-derived human mesenchymal stem cells

Janicki

Boeuf²,

Steck³

et al. 2011

eCM

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Cartilage‐like gene expression in differentiated human stem cell spheroids: A comparison of bone marrow–derived and adipose tissue–derived stromal cells

Cited by 417 publications

References 64 publications

Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice

Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice

Monolayer cell expansion conditions affect the chondrogenic potential of adipose‐derived stem cells

Prediction of in vivo bone forming potency of bone marrow-derived human mesenchymal stem cells

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