Basic Techniques in Human Mesenchymal Stem Cell Cultures: Differentiation into Osteogenic and Adipogenic Lineages, Genetic Perturbations, and Phenotypic Analyses

Bruedigam, Claudia; Driel, Marjolein van; Koedam, Marijke; Peppel, Jeroen van de; Eerden, Bram C. J. van der; Eijken, Marco; Leeuwen, Johannes P.T.M. van

doi:10.1002/9780470151808.sc01h03s17

Cited by 48 publications

(71 citation statements)

References 14 publications

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“…ALP activity is a common marker for osteogenic differentiation [30]. In vitro , during late osteoblast differentiation, ALP dephosphorylates substrates supplied in the media, releasing inorganic phosphate available for the mineralization of the matrix, and its activity rapidly increases during mineralization until a peak level and then subsequently decreases.…”

Section: Resultsmentioning

confidence: 99%

Silk ionomers for encapsulation and differentiation of human MSCs

Calabrese

Kaplan

2012

Biomaterials

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The response of human bone marrow derived human mesenchymal stem cells (hMSCs) encapsulated in silk ionomer hydrogels was studied. Silk aqueous solutions with silk-poly-L-lysine or silk-poly-L-glutamate were formed into hydrogels via ultrasonication in situ with different net charges. hMSCs were encapsulated within the hydrogels and the impact of matrix charge was assessed over weeks in osteogenic, adipogenic and maintenance growth media. These modified silk charged polymers supported cell viability and proliferative potential, and the hMSCs were able to differentiate toward osteogenic or adipogenic lineages in the corresponding differentiation media. The silk/silk-poly-L-lysine hydrogels exhibited a positive effect on selective osteogenesis of hMSCs, inducing differentiation toward an osteogenic lineage even in the absence of osteogenic supplements, while also inhibiting adipogenesis. In contrast, silk/silk fibroin-poly-L-glutamate hydrogels supported both osteogenic and adipogenic differentiation of hMSCs when cultured under induction conditions. The results demonstrate the potential utility of silk-based ionomers in gel formats for hMSCs encapsulation and for directing hMSCs long term functional differentiation toward specific lineages.

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Section: Resultsmentioning

confidence: 99%

Silk ionomers for encapsulation and differentiation of human MSCs

Calabrese

Kaplan

2012

Biomaterials

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“…After centrifugation at 1500 × g for 5 min, the protein content was determined using the DC protein assay kit BioRad. The mineralization content was quantified as previously described in the work of Bruedigam et al [40]. The results are shown as mM Ca 2+ /mg protein.…”

Section: Methodsmentioning

confidence: 99%

Dexamethasone in osteogenic medium strongly induces adipocyte differentiation of mouse bone marrow stromal cells and increases osteoblast differentiation

Ghali¹,

Broux²,

Falgayrac³

et al. 2015

BMC Cell Biol

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BackgroundOsteoblasts and adipocytes share a common mesenchymal stem cell origin. Therefore, it has been suggested that the accumulation of marrow adipocytes observed in bone loss is caused by a shift in the commitment of mesenchymal stem cells from the osteogenic pathway to the adipogenic pathway. Supporting this hypothesis the competition between adipogenic and osteogenic lineages was widely demonstrated on partially homogeneous cell populations. However, some data from mouse models showed the existence of an independent relationship between bone mineral content and bone marrow adiposity. Therefore, the combination of adipogenesis and osteogenesis in primary culture would be helpful to determine if this competition would be observed on a whole bone marrow stromal cell population in a culture medium allowing both lineages.In this aim, mouse bone marrow stromal cells were cultured in a standard osteogenic medium added with different concentrations of Dexamethasone, known to be an important regulator of mesenchymal progenitor cell differentiation.ResultsGene expression of osteoblast and adipocyte markers, biochemical and physical analyses demonstrated the presence of both cell types when Dexamethasone was used at 100 nM. Overall, our data showed that in this co-differentiation medium both differentiation lineages were enhanced compared to classical adipogenic or osteogenic culture medium. This suggests that in this model, adipocyte phenotype does not seem to increase at the expense of the osteoblast lineage.ConclusionThis model appears to be a promising tool to study osteoblast and adipocyte differentiation capabilities and the interactions between these two processes.

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“…PPAR-␥ is expressed in bone marrow stromal cells, adipocytes, osteoblasts, and osteoclasts (13)(14)(15)(16). Although the exact causative factors responsible for the effects of TZDs on bone are not certain, a number of mechanisms have been proposed, including an increase in adipocyte formation at the expense of osteoblast production (17), promotion of osteoclast differentiation and action (18 -20), and stimulation of osteoblast cell apoptosis (21). Other potential actions of TZDs include effects on adipokines and inflammatory cytokines (22)(23)(24), activation of the Wnt signaling pathway (25,26), changes in energy metabolism affecting the skeleton (27), prolonged hyperglycemia (28 -34), inhibition of aromatase, and decreased estrogen synthesis (35).…”

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confidence: 99%