Embryonic rat vascular smooth muscle cells revisited - a model for neonatal, neointimal SMC or differentiated vascular stem cells?

Kennedy, Eimear; Hakimjavadi, Roya; Greene, Chris; Mooney, Ciaran James; Fitzpatrick, Emma; Collins, Laura; Loscher, Christine E.; Guha, Shaunta; Morrow, David; Redmond, Eileen M.; Cahill, Paul A.

doi:10.1186/2045-824x-6-6

Cited by 27 publications

(21 citation statements)

References 46 publications

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“…Consistent with our observations, a recent report revealed that thoracic aortic SMCs of SD rats are immunocytochemically Sox10 + and SM-MHC − , while expressing SMA in both SCGM and RGM (Kennedy et al 2014). Moreover, the expression of SMC markers such as αSMA and calponin is enhanced following the culture of these cells for 7 days in RGM in the presence of TGF-β1 and PDGF (Kennedy et al 2014). These cells were capable of differentiation to adipocytes and osteoblasts (Kennedy et al 2014).…”

Section: Origins Of Mvscssupporting

confidence: 93%

“…Moreover, the expression of SMC markers such as αSMA and calponin is enhanced following the culture of these cells for 7 days in RGM in the presence of TGF-β1 and PDGF (Kennedy et al 2014). These cells were capable of differentiation to adipocytes and osteoblasts (Kennedy et al 2014). This independent study further supports the notion that the primary culture of RASMCs is a process of differentiation of MVSCs.…”

Section: Origins Of Mvscsmentioning

confidence: 96%

“…Given that SMC populations of the thoracic aorta and carotid arteries arise from distinct sources of progenitors during development (Majesky 2007), the MVSCs from aortal and carotid arteries might differ, despite sharing some common features. Of note, a recent report has shown that the synthetic VSMC lines of A10 and A7r5 cells, originally derived from the thoracic aorta of embryonic BD1X rats, express not only the neural stem cell markers of Sox10 and S100β but also Sca-1, c-Kit and Flit-1 and have the potential to differentiate into adipocytes (Kennedy et al 2014). On the other hand, mesenchymal stem-cell-like cells that express CD29 and CD44 have been isolated from bovine aortic media and have multilineage potential for SMC, osteogenic and chondrogenic differentiation but importantly, not for adipogenic differentiation (Tintut et al 2003).…”

Section: Origins Of Mvscsmentioning

confidence: 98%

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Inhibitory role of reactive oxygen species in the differentiation of multipotent vascular stem cells into vascular smooth muscle cells in rats: a novel aspect of traditional culture of rat aortic smooth muscle cells

Song¹,

Wang²,

Wu³

et al. 2015

Cell Tissue Res

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Proliferative or synthetic vascular smooth muscle cells (VSMCs) are widely accepted to be mainly derived from the dedifferentiation or phenotypic modulation of mature contractile VSMCs, i.e., a phenotype switch from a normally quiescent and contractile type into a proliferative or synthetic form. However, this theory has been challenged by recent evidence that synthetic VSMCs predominantly originate instead from media-derived multipotent vascular stem cells (MVSCs). To test these hypotheses further, we re-examine whether the conventional rat aortic SMC (RASMC) culture involves the VSMC differentiation of MVSCs or the dedifferentiation of mature VSMCs and the potential mechanism for controlling the synthetic phenotype of RASMCs. We enzymatically isolated RASMCs and cultured the cells in both a regular growth medium (RGM) and a stem cell growth medium (SCGM). Regardless of culture conditions, only a small portion of freshly isolated RASMCs attaches, survives and grows slowly during the first 7 days of primary culture, while expressing both SMC- and MVSC-specific markers. RGM-cultured cells undergo a process of synthetic SMC differentiation, whereas SCGM-cultured cells can be differentiated into not only synthetic SMCs but also other somatic cells. Notably, compared with the RGM-cultured differentiated RASMCs, the SCGM-cultured undifferentiated cells exhibit the phenotype of MVSCs and generate greater amounts of reactive oxygen species (ROS) that act as a negative regulator of differentiation into synthetic VSMCs. Knockdown of phospholipase A2, group 7 (Pla2g7) suppresses ROS formation in the MVSCs while enhancing SMC differentiation of MVSCs. These results suggest that cultured synthetic VSMCs can be derived from the SMC differentiation of MVSCs with ROS as a negative regulator.

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Section: Origins Of Mvscssupporting

confidence: 93%

Section: Origins Of Mvscsmentioning

confidence: 96%

Section: Origins Of Mvscsmentioning

confidence: 98%

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Inhibitory role of reactive oxygen species in the differentiation of multipotent vascular stem cells into vascular smooth muscle cells in rats: a novel aspect of traditional culture of rat aortic smooth muscle cells

Song¹,

Wang²,

Wu³

et al. 2015

Cell Tissue Res

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show abstract

“…Interestingly, MVSCs were also labeled in the common carotid artery in the Wnt1-Cre reporter mice [35], suggesting that MVSCs could be involved in neointima formation in the vein or artery grafts. Because MVSCs are SMMHC negative [22, 35], while we found that the GFP-positive cells in common carotid artery in Wnt1-Cre + transgenic mice were SMMHC positive (Fig. 4c), suggest that those GFP/SMMHC positive SMCs are precursors that contribute to neointima cells.…”

Section: Discussionmentioning

confidence: 77%

Smooth muscle cells from the anastomosed artery are the major precursors for neointima formation in both artery and vein grafts

Liang

Wang

et al. 2014

Basic Res Cardiol

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Accumulation of smooth muscle cells (SMC) results in neointima formation in injured vessels. Two graft models consisting of vein and artery grafts were created by anastomosing common carotid arteries to donor vessels. To identify the origin of the neointima cells from anastomosed arteries, we use Wnt1-Cre/reporter mice to label and track SMCs in the common carotid artery. The contribution of SMCs in the neighboring arteries to neointima formation was studied. On evaluating the artery grafts after 1 month, >90 % of the labeled neointima cells were found to have originated from the anastomosing host arteries. Most of the neointima cells were also smooth muscle α-actin positive (SMA-α+) and expressed the smooth muscle myosin heavy chain (SMMHC), the SMC terminal differentiation marker. In vein grafts, about 60 % SMA-α-positive cells were from anastomosing arteries. Bone marrow cells did not contribute to neointima SMCs in vein grafts, but did co-stain with markers of inflammatory cells. Wnt1 expression was not detected in the neointima cells in the vein or artery grafts, or the injured femoral arteries. Neointima SMCs showed the synthetic phenotype and were positively labeled with BrdU in vitro and in vivo. Treatment with the IGF-1 receptor inhibitor suppressed SMC proliferation and neointima formation in vein grafts. Our results indicate that SMCs from the neighboring artery are predominantly present in the neointima formed in both vein and artery grafts and that Wnt1-Cre mice can be used to explore the role of SMCs originating from neighboring vessels in vascular remodeling.

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“…In addition to cancer cells, we also monitored the effects of seeding density on the proliferation of TRPC1 -silenced A7r5 cells which has a significance in examining the vascular contractile and proliferative phenotypes in vitro [36]. It is known that vascular smooth muscle cells have the ability of plasticity with a wide range of phenotypes besides their primary contractile phenotype [37].…”

Section: Discussionmentioning

confidence: 99%

Effects of cell seeding density on real-time monitoring of anti-proliferative effects of transient gene silencing

Selli

Erac

Tosun

2016

J of Biol Res-Thessaloniki

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BackgroundReal-time cellular analysis systems enable impedance-based label-free and dynamic monitoring of various cellular events such as proliferation. In this study, we describe the effects of initial cell seeding density on the anti-proliferative effects of transient gene silencing monitored via real-time cellular analysis. We monitored the real-time changes in proliferation of Huh7 hepatocellular carcinoma and A7r5 vascular smooth muscle cells with different initial seeding densities following transient receptor potential canonical 1 (TRPC1) silencing using xCELLigence system. Huh7 and A7r5 cells were seeded on E-plate 96 at 10,000, 5000, 1250 and 5000, 2500 cells well−1, respectively, following silencing vector transfection. The inhibitory effects of transient silencing on cell proliferation monitored every 30 min for 72 h.ResultsTRPC1 silencing did not inhibit the proliferation rates of Huh7 cells at 10,000 cells well−1 seeding density. However, a significant anti-proliferative effect was observed at 1250 cells well−1 density at each time point throughout 72 h. Furthermore, significant inhibitory effects on A7r5 proliferation were observed at both 5000 and 2500 cells well−1 for 72 h.ConclusionsData suggest that the effects of transient silencing on cell proliferation differ depending on the initial cell seeding density. While high seeding densities mask the significant changes in proliferation, the inhibitory effects of silencing become apparent at lower seeding densities as the entry into log phase is delayed. Using the optimal initial seeding density is crucial when studying the effects of transient gene silencing. In addition, the results suggest that TRPC1 may contribute to proliferation and phenotypic switching of vascular smooth muscle cells.

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Embryonic rat vascular smooth muscle cells revisited - a model for neonatal, neointimal SMC or differentiated vascular stem cells?

Cited by 27 publications

References 46 publications

Inhibitory role of reactive oxygen species in the differentiation of multipotent vascular stem cells into vascular smooth muscle cells in rats: a novel aspect of traditional culture of rat aortic smooth muscle cells

Inhibitory role of reactive oxygen species in the differentiation of multipotent vascular stem cells into vascular smooth muscle cells in rats: a novel aspect of traditional culture of rat aortic smooth muscle cells

Smooth muscle cells from the anastomosed artery are the major precursors for neointima formation in both artery and vein grafts

Effects of cell seeding density on real-time monitoring of anti-proliferative effects of transient gene silencing

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