Effect of fibronectin, FGF-2, and BMP4 in the stemness maintenance of BMSCs and the metabolic and proteomic cues involved

Abstract

Background Growing evidence suggests that the pluripotent state of mesenchymal stem cells (MSCs) relies on specific local microenvironmental cues such as adhesion molecules and growth factors. Fibronectin (FN), fibroblast growth factor 2 (FGF2), and bone morphogenetic protein 4 (BMP4) are the key players in the regulation of stemness and lineage commitment of MSCs. Therefore, this study was designed to investigate the pluripotency and multilineage differentiation of bone marrow-derived MSCs (BMSCs) with the introduction of FN, FGF-2, and BMP4 and to identify the metabolic and proteomic cues involved in stemness maintenance. Methods To elucidate the stemness of BMSCs when treated with FN, FGF-2, and BMP4, the pluripotency markers of OCT4, SOX2, and c-MYC in BMSCs were monitored by real-time PCR and/or western blot. The nuclear translocation of OCT4, SOX2, and c-MYC was investigated by immunofluorescence staining. Multilineage differentiation of the treated BMSCs was determined by relevant differentiation markers. To identify the molecular signatures of BMSC stemness, gas chromatography-mass spectrometry (GC-MS), liquid chromatography-tandem mass spectrometry (LC-MS/MS), and bioinformatics analysis were utilized to determine the metabolite and protein profiles associated with stem cell maintenance. Results Our results demonstrated that the expression of stemness markers decreased with BMSC passaging, and the manipulation of the microenvironment with fibronectin and growth factors (FGF2 and BMP4) can significantly improve BMSC stemness. Of note, we revealed 7 differentially expressed metabolites, the target genes of these metabolites may have important implications in the maintenance of BMSCs through their effects on metabolic activity, energy production, and potentially protein production. We also identified 21 differentially abundant proteins, which involved in multiple pathways, including metabolic, autophagy-related, and signaling pathways regulating the pluripotency of stem cells. Additionally, bioinformatics analysis comfirned the correlation between metabolic and proteomic profiling, suggesting that the importance of metabolism and proteome networks and their reciprocal communication in the preservation of stemness. Conclusions These results indicate that the culture environment supplemented with the culture cocktail (FN, FGF2, and BMP4) plays an essential role in shaping the pluripotent state of BMSCs. Both the metabolism and proteome networks are involved in this process and the modulation of cell-fate decision making. All these findings may contribute to the application of MSCs for regenerative medicine.

Umbilical Cord Mesenchymal Stem Cell Therapy for Regenerative Treatment of Rheumatoid Arthritis: Opportunities and Challenges

“…UC-MSCs secrete a range of growth factors, such as hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), stromal cell-derived factor-1 (SDF-1), keratinocyte growth factor (KGF), fibroblast growth factor (FGF), and insulin-like growth factor-1 (IGF-1), to facilitate proliferation and tissue damage repair. 45 , 46 MSCs have the potential to be pluripotent and capable of giving rise to a variety of cell types. The properties of various factors, growth factors, and extracellular vesicles can be transmitted.…”

Construction of a myocardial patch with mesenchymal stem cells and poly(CL-co-TOSUO)/collagen scaffolds for myocardial infarction repair by coaxial electrospinning

“…[31][32][33] Many studies have reported that MSCs could greatly regulate inflammation and improve MI cardiac angiogenesis, and these regeneration effects were greatly related to the stem cell properties. [34][35][36][37] In this study, after 5-day dynamic cultivation in vitro, MSCs maintained high expressions of typical stem cell markers CD29 and CD90 on the PCT/collagen scaffold. Moreover, in vitro results showed that the PCT/collagen scaffolds were beneficial for MSCs adhesion and proliferation, and they could provide physical protection for MSCs in infarct area.…”

Bmp5 Mutation Alters miRNA Expression During Embryonic External Ear Development

“…BMP family genes may play a regulatory role in pluripotency and multiline differentiation of bone marrow-derived mesenchymal stem cells and can participate in dry maintenance metabolism and proteomics. 14 The expression of miRNAs related to the MAPK signaling pathway, PI3K-Akt signaling pathway, focal adhesion signaling pathway, Rap1 signaling pathway, metabolic pathways, and pathways related to cancer, apoptosis, and axon guidance increased during the development of E15.5 and E17.5 embryos of Bmp5 short ear mice. The growth factor signal mediated by receptor tyrosine kinases is important for the normal formation and development of the NC.…”

“…BMP family genes may play a regulatory role in pluripotency and multiline differentiation of bone marrow-derived mesenchymal stem cells and can participate in dry maintenance metabolism and proteomics. 14…”