Implication of NOD1 and NOD2 for the Differentiation of Multipotent Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

Abstract

Toll-like receptors (TLRs) and Nod-like receptors (NLRs) are known to trigger an innate immune response against microbial infection. Although studies suggest that activation of TLRs modulate the function of mesenchymal stem cells (MSCs), little is known about the role of NLRs on the MSC function. In this study, we investigated whether NOD1 and NOD2 regulate the functions of human umbilical cord blood-derived MSCs (hUCB-MSCs). The genes of TLR2, TLR4, NOD1, and NOD2 were expressed in hUCB-MSCs. Stimulation with each agonist (Pam3CSK4 for TLR2, LPS for TLR4, Tri-DAP for NOD1, and MDP for NOD2) led to IL-8 production in hUCB-MSC, suggesting the expressed receptors are functional in hUCB-MSC. CCK-8 assay revealed that none of agonist influenced proliferation of hUCB-MSCs. We next examined whether TLR and NLR agonists affect osteogenic-, adipogenic-, and chondrogenic differentiation of hUCB-MSCs. Pam3CSK4 and Tri-DAP strongly enhanced osteogenic differentiation and ERK phosphorylation in hUCB-MSCs, and LPS and MDP also slightly did. Treatment of U0126 (MEK1/2 inhibitor) restored osteogenic differentiation enhanced by Pam3CSK4. Tri-DAP and MDP inhibited adipogenic differentiation of hUCB-MSCs, but Pam3CSK4 and LPS did not. On chondrogenic differentiation, all TLR and NLR agonists could promote chondrogenesis of hUCB-MSCs with difference in the ability. Our findings suggest that NOD1 and NOD2 as well as TLRs are involved in regulating the differentiation of MSCs.

Nod1 Activation by Bacterial iE-DAP Induces Maternal–Fetal Inflammation and Preterm Labor

“…Based on these studies, and because we did not find any influx of maternal immune cells into the placental-uterine units after iE-DAP administration, our findings support a specific fetal immune response, which can arise from either maternal cytokines trafficking to and stimulating the fetal immune system or from the movement of iE-DAP across the placental to the fetus, again stimulating the fetal immune system. Indeed, fetal PRRs such as the TLRs have been implicated in the fetal inflammatory response syndrome (56), and Nod1 and Nod2 may be involved in regulating the differentiation of human umbilical cord blood mesenchymal stem cells (57). In summary, we have found that term human trophoblast cells express functional Nod1, but not Nod2.…”

Mesenchymal Stromal Cells in Transplantation Rejection and Tolerance

“…MSCs are known to express a number of PRRs, including TLR1-9 (Pevsner-Fischer et al 2007;Tomchuck et al 2008;Opitz et al 2009;Romieu-Mourez et al 2009), NOD receptors (Kim et al 2010;Sioud et al 2010), and RAGE (Kume et al 2005). These receptors are functionally active on MSCs, and binding to their respective ligands leads to alterations in MSC functions.…”

Stimulative effects ofPolygonum amplexicaulevar.sinenseon osteoblastic MC3T3-E1 cells

“…After washing twice with phosphate-buffered saline (PBS), the cells were treated with various concentrations of PAF ethanol extracts for 1-7 days, and cell proliferation was evaluated by Cell Count Kit-8 assay (CCK-8; Beyotime Inst Biotech., China) according to the manufacturer's instructions (Kim et al, 2010). In brief, 10 μL CCK-8 was added to each well (100 μL medium).…”