Growth inhibition and differentiation of cultured smooth muscle cells depend on cellular crossbridges across the tubular lumen of type I collagen matrix honeycombs

Suzuki, Takaaki; Ishii, Itsuko; Kotani, Akio; Masuda, Michi; Hirata, Kaori; Ueda, Madoka; Ogata, Takahiro; Sakai, Takanori; Ariyoshi, Noritaka; Kitada, Mitsukazu

doi:10.1016/j.mvr.2008.08.006

Cited by 6 publications

(5 citation statements)

References 18 publications

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“…SMCs in honeycombs are attached across the lumens of the honeycombs by a few adhesion points and have a more spindly shape than do SMCs cultured on plastic plates. We have also reported that formation of cellular crossbridges in honeycombs is important for changes that occur in SMCs grown in honeycombs [6]. These results suggest that relationships exist between the structural rearrangement of the cytoskeleton and specific features of SMCs cultured in honeycombs.…”

Section: Introductionmentioning

confidence: 53%

“…SMCs cultured in honeycombs also stopped proliferating and did not respond to PDGF-BB anymore [5]. Additionally, SMCs cultured in honeycombs expressed caldesmon heavy chain (a marker protein of mature differentiated SMCs) [5,6]. Although expression levels of -actin and MHC (a marker of differentiated SMCs) in SMCs cultured on plates and in honeycombs were almost identical, SMCs in honeycombs might be able to achieve contractility.…”

Section: Discussionmentioning

confidence: 99%

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Degradation of Filamin Induces Contraction of Vascular Smooth Muscle Cells in Type-I Collagen Matrix Honeycombs

Uchida

Ishii

Hirata

et al. 2011

Cell Physiol Biochem

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Background: Dedifferentiated rabbit vascular smooth muscle cells (SMCs) exhibit similar features to differentiated SMCs when cultured in three-dimensional matrices of type-I collagen called “honeycombs,” but the mechanism is unknown. The role of filamin, an actin-binding protein that links actin filaments in SMCs, was investigated. Methods: Filamin and other related proteins were detected by western blot analysis and immunofluorescence staining. Honeycomb size was measured to confirm the contraction of SMCs. Results: Full-length filamin was expressed in subconfluent SMCs cultured on plates; however, degradation of filamin, which might be regulated by calpain, was observed in confluent SMCs cultured on plates and in honeycombs. While filamin was co-localized with β-actin in subconfluent SMCs grown on plates, filamin was detected in the cytoplasm in SMCs cultured in honeycombs, and degraded filamin was mainly detected in the cytoplasmic fraction of these cells. In addition, β-actin expression was low in the cytoskeletal fraction of SMCs cultured in honeycombs compared with cells cultured on plates, and the size of the honeycombs used for culturing SMCs was significantly reduced. Conclusion: These data suggest that degradation of filamin in SMCs cultured in honeycombs induces structural weakness of β-non-muscle actin filaments, thereby permitting SMCs in honeycombs to achieve contractility.

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Section: Introductionmentioning

confidence: 53%

Section: Discussionmentioning

confidence: 99%

Degradation of Filamin Induces Contraction of Vascular Smooth Muscle Cells in Type-I Collagen Matrix Honeycombs

Uchida

Ishii

Hirata

et al. 2011

Cell Physiol Biochem

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“…VSMCs can be cultured in honeycombs for approximately 3 months with medium change. However, when rabbit VSMCs are cultured in honeycombs with larger pore size (100-500 μm) (Figure 3), they proliferate for the first few days, but then stop proliferating and cell number does not change [6]. The reason why cell number increases only at the beginning of culture is as follows: in honeycombs with a smaller pore (≤200 μm), VSMCs form cross-bridges independently of each other, but a large number of connected cells form cross-bridges together at the wall of honeycombs with larger pores (100-500 μm) after the initial increase in cell number at the beginning of culture.…”

Section: Proliferative Inhibition Of Vsmcs Cultured In Honeycombsmentioning

confidence: 99%

“…The depletion of caldesmon facilitates the formation of podosomes/invadopodia and cell invasion [19]. As VSMCs cultured in honeycombs for 14 days express h-CaD [4,6], these cells could be classified to the contractile phenotype.…”

Section: Differentiation Of Vsmcs Cultured In Honeycombsmentioning

confidence: 99%

Three-Dimensional “Honeycomb” Culture System that Helps to Maintain the Contractile Phenotype of Vascular Smooth Muscle Cells

Ishii¹,

Uchida²

2015

Muscle Cell and Tissue

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Vascular smooth muscle cells (VSMCs) in the normal aorta are described as having a contractile phenotype because they can contract and do not proliferate. VSMCs in pathological conditions such as atherosclerosis and restenosis can proliferate and migrate, but lose their ability to contract, which is referred to as a synthetic phenotype. VSMCs show plasticity by changing their phenotype according to the surrounding environment. When VSMCs are cultured on a plastic plate, which is a normal two-dimensional culture system, they display the synthetic phenotype because they proliferate and migrate without contraction. Recently, we successfully cultured VSMCs that display features similar to the contractile phenotype, using type I collagen three-dimensional matrices, "honeycombs," in the presence of abundant fetal bovine serum albumin. VSMCs cultured in honeycombs stop proliferating and can contract. The honeycomb culture system can maintain VSMCs in the contractile phenotype for a long period of time. In this chapter, we show the method of this new culture system and the characteristics of VSMCs in honeycombs. It is expected that the use of this culture system will generate new information on the characteristics of VSMCs.

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“…The disadvantage of hydrogels is the lack of vessel structure and mechanical strength, hence additional support is required. Collagen can also be formed into honeycomb-like scaffolds of varying pore sizes that have been found to induce proper VSMC phenotype and differentiation markers better than collagen-coated films [73].…”

Section: Extracellular Matrix-based Scaffoldsmentioning

confidence: 99%

Tissue Engineering a Small Diameter Vessel Substitute: Engineering Constructs with Select Biomaterials and Cells

McBane

Sharifpoor²,

Labow³

et al. 2012

CVP

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Cardiovascular disease (CVD) is a leading cause of death and hospitalization worldwide. The need for small caliber vessels ( < 6mm) to treat CVD patients has grown; however the availability of autologous vessels in cardiac and peripheral bypass candidates is limited. The search for an alternative vessel source is widespread with both natural and synthetic tissue engineered materials being investigated as scaffolds. Despite decades of exhaustive studies with decellularized extracellular matrices (ECM) and synthetic graft materials, the field remains in search of a commercially viable biomaterial construct substitute. While the previous materials have been assessed by evaluating their compatibility with fibroblasts, smooth muscle cells and endothelial cells, current materials are being conceived based on their interactions with stem cells, progenitor cells and monocytes, as the latter may hold the key to repair and regeneration. The graft's ability to recruit and maintain these cells has become a major research focus. The successful tissue engineering of a small caliber vessel graft requires the use of optimal material chemistry and biological function to promote cell recruitment into the graft while maintaining each functional phenotype during vessel tissue maturation. The discussion of these significant research challenges constitutes the focus of this review.

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Growth inhibition and differentiation of cultured smooth muscle cells depend on cellular crossbridges across the tubular lumen of type I collagen matrix honeycombs

Cited by 6 publications

References 18 publications

Degradation of Filamin Induces Contraction of Vascular Smooth Muscle Cells in Type-I Collagen Matrix Honeycombs

Degradation of Filamin Induces Contraction of Vascular Smooth Muscle Cells in Type-I Collagen Matrix Honeycombs

Three-Dimensional “Honeycomb” Culture System that Helps to Maintain the Contractile Phenotype of Vascular Smooth Muscle Cells

Tissue Engineering a Small Diameter Vessel Substitute: Engineering Constructs with Select Biomaterials and Cells

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