Smooth muscle cells differentiated from mesenchymal stem cells are regulated by microRNAs and suitable for vascular tissue grafts

Abstract

Tissue-engineered vascular grafts with long-term patency are greatly needed in the clinical settings, and smooth muscle cells (SMCs) are a critical graft component. Human mesenchymal stem cells (MSCs) are used for generating SMCs, and understanding the underlying regulatory mechanisms of the MSC-to-SMC differentiation process could improve SMC generation in the clinic. Here, we found that in response to stimulation of transforming growth factor-β1 (TGFβ1), human umbilical cord–derived MSCs abundantly express the SMC markers α-smooth muscle actin (αSMA), smooth muscle protein 22 (SM22), calponin, and smooth muscle myosin heavy chain (SMMHC) at both gene and protein levels. Functionally, MSC-derived SMCs displayed contracting capacity in vitro and supported vascular structure formation in the Matrigel plug assay in vivo. More importantly, SMCs differentiated from human MSCs could migrate into decellularized mouse aorta and give rise to the smooth muscle layer of vascular grafts, indicating the potential of utilizing human MSC-derived SMCs to generate vascular grafts. Of note, microRNA (miR) array analysis and TaqMan microRNA assays identified miR-503 and miR-222-5p as potential regulators of MSC differentiation into SMCs at early time points. Mechanistically, miR-503 promoted SMC differentiation by directly targeting SMAD7, a suppressor of SMAD-related, TGFβ1-mediated signaling pathways. Moreover, miR-503 expression was SMAD4-dependent. SMAD4 was enriched at the miR-503 promoter. Furthermore, miR-222-5p inhibited SMC differentiation by targeting and down-regulating ROCK2 and αSMA. In conclusion, MSC differentiation into SMCs is regulated by miR-503 and miR-222-5p and yields functional SMCs for use in vascular grafts.

Heterotypic Scaffold Design Orchestrates Primary Cell Organization and Phenotypes in Cocultured Small Diameter Vascular Grafts

“…The last key consideration point for aiming at engineering of the tunica media is to realize the SMC phenotype switch to the contractile phenotype at the proper stage during their maturation process on the scaffold. Generally, differentiation of MSCs into contractile vSMCs is accomplished by the addition of biochemical factors associated with differentiation, such as TGF‐β1 or platelet‐derived growth factor subunit β 22d,29,31. Interestingly, we observed that by expanding bone marrow‐derived MSCs in a culture plate, supplemented with the proliferation‐associated basic fibroblast growth factor (bFGF), also an induction of differentiation was observed after reaching confluency.…”

Directional Topography Influences Adipose Mesenchymal Stromal Cell Plasticity: Prospects for Tissue Engineering and Fibrosis

“…The generation of tissue-engineered blood vessels (TEBV) is facilitated by the use of mesenchymal stem cells (MSC) as precursors for vascular smooth muscle cells (SMC), i.e., the media layer of TEBV [1–5]. These highly plastic MSC, originating from the mesodermal embryonic tissue and present in all connective tissues of the adult human body, are relatively easy to isolate, cultivate, and characterize [6, 7].…”

Glycoprotein M6B Interacts with TβRI to Activate TGF-β-Smad2/3 Signaling and Promote Smooth Muscle Cell Differentiation

“…DeltaEF1, lncRNA growth arrest‐specific 5 (GAS5), and brain cytoplasmic RNA 1 (BC1) can specifically bind to Smad3 protein and reduce Smad3 translocation to the nucleus or bind to TGF‐β‐responsive SMC gene promoters . In addition, miRNA‐503 induces SMC differentiation through TGF‐β signaling by inhibition of Smad7 transcription . Although extensive studies have focused on TGF‐β1‐induced SMC differentiation, the molecular mechanisms controlling the TGF‐β signaling during SMC differentiation are still not fully known.…”