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Toll-like receptor triggering in cord blood mesenchymal stem cells.

van den Berk LC, Jansen BJ, Siebers-Vermeulen KG, Netea MG, Latuhihin T, Bergevoet S, Raymakers RA, Kögler G, Figdor CC, Adema GJ, Torensma R - J. Cell. Mol. Med. (2009)

Bottom Line: Recently, the antagonizing effect on the differentiation of mesenchymal stem cells (MSCs) by toll-like receptor (TLR) ligands, was described.In summary, TLR4 and 5 signalling in USSCs is slow and results in the up-regulation of a restricted number of pro-inflammatory cytokines and enhanced osteoblastic differentiation.Apparently, the outcome of TLR signalling depends on the cell type that expresses them.

View Article: PubMed Central - PubMed

Affiliation: Department of Tumor Immunology, Nijmegen Centre for Molecular Life Sciences, Geert Grooteplein, Nijmegen, The Netherlands.

ABSTRACT
Recently, the antagonizing effect on the differentiation of mesenchymal stem cells (MSCs) by toll-like receptor (TLR) ligands, was described. Our study shows that on more primitive cord blood derived MSCs, the expression of TLRs and ligand-induced triggering differs from that of bone marrow derived MSCs. At the RNA level, cord blood MSCs (unrestricted somatic stem cells; USSCs) express low levels of TLR1,3,5,9 and high levels of TLR4 and TLR6. At the protein level expression of TLR5 and very low expression of TLR4 was observed. NF-kappaB translocation studies revealed that both TLR4 and TLR5 are functional, although signalling kinetics induced by the individual ligands differed. Stimulation of USSCs with either lipopolysaccharide (LPS) or flagellin resulted in a marked increase of interleukin (IL)-6 and/or IL-8 production although levels differed significantly between both stimuli. Interestingly, tumour necrosis factor (TNF)-alpha was undetectable after TLR stimulation, which appeared to be due to an inactivated TNF-alpha promoter in USSCs. Moreover, osteoblastic differentiation was enhanced after triggering USSCs with LPS and flagellin. In summary, TLR4 and 5 signalling in USSCs is slow and results in the up-regulation of a restricted number of pro-inflammatory cytokines and enhanced osteoblastic differentiation. Apparently, the outcome of TLR signalling depends on the cell type that expresses them.

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LPS and flagellin promote osteogenic differentiation of USSCs. USSCs were differentiated towards the osteogenic lineage for 6 to 14 days in the absence (–) or presence of TLR agonists (LPS, flagellin or polyI: C). ALP and ARS stainings of 6, 7 and 8 days differentiated USSCs are shown.
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fig07: LPS and flagellin promote osteogenic differentiation of USSCs. USSCs were differentiated towards the osteogenic lineage for 6 to 14 days in the absence (–) or presence of TLR agonists (LPS, flagellin or polyI: C). ALP and ARS stainings of 6, 7 and 8 days differentiated USSCs are shown.

Mentions: USSCs have the ability to differentiate into several lineages, such as osteoblasts, chondrocytes, adipocytes, haematopoietic cells, liver and neural and heart tissue [19, 20]. The most robust differentiation pathway is the osteoblastic lineage. Therefore, TLR agonists were added to the differentiation media to assess whether TLR agonists affect the osteogenic differentiation of USSCs. Cells were differentiated for 6 to 14 days and osteogenic differentiation was detected by ALP or ARS staining. LPS and flagellin strongly stimulated the initial phase of osteogenic differentiation of USSCs as compared to the normal control. As a negative control polyI: C was used (Fig. 7), since this TLR agonist had no effect on NF-κB signalling, proliferation and immunomodulatory properties. After day 11 of differentiation no difference in osteogenic differentiation was observed (data not shown). Additionally, USSCs cultured in normal USSC medium stained negative for both ALP and ARS (data not shown). Identical results were found for both USSC lines.


Toll-like receptor triggering in cord blood mesenchymal stem cells.

van den Berk LC, Jansen BJ, Siebers-Vermeulen KG, Netea MG, Latuhihin T, Bergevoet S, Raymakers RA, Kögler G, Figdor CC, Adema GJ, Torensma R - J. Cell. Mol. Med. (2009)

LPS and flagellin promote osteogenic differentiation of USSCs. USSCs were differentiated towards the osteogenic lineage for 6 to 14 days in the absence (–) or presence of TLR agonists (LPS, flagellin or polyI: C). ALP and ARS stainings of 6, 7 and 8 days differentiated USSCs are shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4516497&req=5

fig07: LPS and flagellin promote osteogenic differentiation of USSCs. USSCs were differentiated towards the osteogenic lineage for 6 to 14 days in the absence (–) or presence of TLR agonists (LPS, flagellin or polyI: C). ALP and ARS stainings of 6, 7 and 8 days differentiated USSCs are shown.
Mentions: USSCs have the ability to differentiate into several lineages, such as osteoblasts, chondrocytes, adipocytes, haematopoietic cells, liver and neural and heart tissue [19, 20]. The most robust differentiation pathway is the osteoblastic lineage. Therefore, TLR agonists were added to the differentiation media to assess whether TLR agonists affect the osteogenic differentiation of USSCs. Cells were differentiated for 6 to 14 days and osteogenic differentiation was detected by ALP or ARS staining. LPS and flagellin strongly stimulated the initial phase of osteogenic differentiation of USSCs as compared to the normal control. As a negative control polyI: C was used (Fig. 7), since this TLR agonist had no effect on NF-κB signalling, proliferation and immunomodulatory properties. After day 11 of differentiation no difference in osteogenic differentiation was observed (data not shown). Additionally, USSCs cultured in normal USSC medium stained negative for both ALP and ARS (data not shown). Identical results were found for both USSC lines.

Bottom Line: Recently, the antagonizing effect on the differentiation of mesenchymal stem cells (MSCs) by toll-like receptor (TLR) ligands, was described.In summary, TLR4 and 5 signalling in USSCs is slow and results in the up-regulation of a restricted number of pro-inflammatory cytokines and enhanced osteoblastic differentiation.Apparently, the outcome of TLR signalling depends on the cell type that expresses them.

View Article: PubMed Central - PubMed

Affiliation: Department of Tumor Immunology, Nijmegen Centre for Molecular Life Sciences, Geert Grooteplein, Nijmegen, The Netherlands.

ABSTRACT
Recently, the antagonizing effect on the differentiation of mesenchymal stem cells (MSCs) by toll-like receptor (TLR) ligands, was described. Our study shows that on more primitive cord blood derived MSCs, the expression of TLRs and ligand-induced triggering differs from that of bone marrow derived MSCs. At the RNA level, cord blood MSCs (unrestricted somatic stem cells; USSCs) express low levels of TLR1,3,5,9 and high levels of TLR4 and TLR6. At the protein level expression of TLR5 and very low expression of TLR4 was observed. NF-kappaB translocation studies revealed that both TLR4 and TLR5 are functional, although signalling kinetics induced by the individual ligands differed. Stimulation of USSCs with either lipopolysaccharide (LPS) or flagellin resulted in a marked increase of interleukin (IL)-6 and/or IL-8 production although levels differed significantly between both stimuli. Interestingly, tumour necrosis factor (TNF)-alpha was undetectable after TLR stimulation, which appeared to be due to an inactivated TNF-alpha promoter in USSCs. Moreover, osteoblastic differentiation was enhanced after triggering USSCs with LPS and flagellin. In summary, TLR4 and 5 signalling in USSCs is slow and results in the up-regulation of a restricted number of pro-inflammatory cytokines and enhanced osteoblastic differentiation. Apparently, the outcome of TLR signalling depends on the cell type that expresses them.

Show MeSH
Related in: MedlinePlus