FGF2 induces RANKL gene expression as well as IL1β regulated MHC class II in human bone marrow-derived mesenchymal progenitor stromal cells

Abstract

ObjectiveHuman bone marrow mesenchymal stromal cells (hBM-MSC) are being applied in tissue regeneration and treatment of autoimmune diseases (AD). Their cellular and immunophenotype depend on isolation and culture conditions which may influence their therapeutic application and reflect their in vivo biological functions. We have further characterised the phenotype induced by fibroblast growth factor 2 (FGF2) on healthy donor hBM-MSC focusing on the osteoimmunological markers osteoprotegerin (OPG), receptor activator of nuclear factor kB (RANK), RANK ligand (RANKL) and HLA-DR and their regulation of expression by the inflammatory cytokines IL1β and IFNγ.MethodsRANK, RANKL, OPG and HLA-DR expression in hBM-MSC expanded under specific culture conditions, were measured by RT-PCR and flow cytometry. MAPKs induction by FGF2, IL1β and IFNγ in hBM-MSC was analysed by immunoblotting and RT-PCR.ResultsIn hBM-MSC, OPG expression is constitutive and FGF2 independent. RANKL expression depends on FGF2 and ERK1/2 activation. IL1β and IFNγ activate ERK1/2 but fail to induce RANKL. Only IL1β induces P38MAPK. The previously described HLA-DR induced by FGF2 through ERK1/2 on hBM-MSC, is suppressed by IL1β through inhibition of CIITA transcription. HLA-DR induced by IFNγ is not affected by IL1β in hBM-MSC, but is suppressed in articular chondrocytes and lung fibroblasts.ConclusionsRANKL expression and IL1β regulated MHC-class II, both induced via activation of the ERK1/2 signalling pathway, are specific for progenitor hBM-MSC expanded in the presence of FGF2. HLA-DR regulated by IL1β and ERK1/2 is observed on hBM-MSC during early expansion without FGF2 suggesting previous in vivo acquisition. Stromal progenitor cells with this phenotype could have an osteoimmunological role during bone regeneration.

Murine Rankl−/− Mesenchymal Stromal Cells Display an Osteogenic Differentiation Defect Improved by a RANKL-Expressing Lentiviral Vector

“…To the best of our knowledge, this hypothesis has not been addressed in detail in literature. In a recent article, the authors could detect RANK expression in human BM‐MSCs only in the presence of FGF2 in the culture medium ; however, this discrepancy with our own result might derive from different conditions of isolation, culture and expansion. Therefore, further evaluating the possibility of a RANKL/RANK axis in MSCs could be highly relevant, considering that the RANKL molecule is a target of anti‐resorptive treatments in humans.…”

Production of oncolytic adenovirus and human mesenchymal stem cells in a single‐use, Vertical‐Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier‐based cell culture processes

“…It is well known that the cellular and immunophenotype of hMSC depend not only on the isolation protocol but also on the culture conditions. 57,58 Our results showed that both bioreactor types were able to maintain the hMSC cellular phenotype; hMSC were negative for hematopoietic CD34 and CD45 markers and displayed high levels of CD44, CD73, CD105, CD90, and CD166 mesenchymal stem markers ( Figures 6A and B). According to the International Society for Cellular Therapy, hMSC are also considered HLA-DR-negative with no/reduced (below 5%) expression of HLA-DR surface molecules.…”

Withdrawn: Geraniin inhibits TNF-α-induced impairments of osteogenesis through NF-κB and p38 MAPK signalling pathways in bone marrow stem cells

“… 27 TNF-α can promote the secretion of Hepatocyte growth factor by BMSCs through p38 MAPK and PI3K/Akt pathways. 28 Thus, NF-κB, ERK and c-Jun-NH2-terminal Kinase (JNK) pathways play an essential role for migration homing and adhesion of BMSCs. 29 …”