Contrasting effect of perlecan on adipogenic and osteogenic differentiation of mesenchymal stem cells <i>in vitro</i>

Nakamura, Ryosuke; Nakamura, Fumio; Fukunaga, Shigeharu

doi:10.1111/asj.12116

Cited by 14 publications

(15 citation statements)

References 29 publications

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“…Bovine kidney perlecan and HS-truncated perlecan (perlecan ΔHS) were prepared as described previously [Nakamura et al, 2014a]. The purity of perlecan was verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, followed by heparitinase-I and/or chondroitinase ABC treatment ( fig.…”

Section: Methodsmentioning

confidence: 99%

“…The purity of perlecan was verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, followed by heparitinase-I and/or chondroitinase ABC treatment ( fig. 1 ) [Nakamura et al, 2014a]. The purified perlecan was substituted with HS but not with chondroitin sulfate chains.…”

Section: Methodsmentioning

confidence: 99%

“…Anti-perlecan antiserum was prepared as described previously [Nakamura et al, 2014a]. The antiserum was generated in a rabbit by immunizing the purified bovine kidney perlecan, which was not treated with any HS-degrading enzymes.…”

Section: Methodsmentioning

confidence: 99%

“…Heparitinase-I could not be detected by CBB staining. The data are adapted from Nakamura et al [2014a].…”

Section: Methodsmentioning

confidence: 99%

“…Salted-out conditioned media were spotted onto nitrocellulose membranes. The membranes were subsequently rinsed with PBS containing 0.05% Tween 20 (T-PBS), blocked with T-PBS containing 2% casein (Wako) and 1% bovine serum albumin (BSA; Wako), and incubated with anti-perlecan antiserum [Nakamura et al, 2014a]. The membranes were washed with T-PBS three times and incubated with horseradish peroxidase-conjugated anti-rabbit immunoglobulin (Ig)G, IgA and IgM (MP Biomedicals, Solon, Ohio, USA).…”

Section: Dot Blot Analysismentioning

confidence: 99%

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Perlecan Diversely Regulates the Migration and Proliferation of Distinct Cell Types in vitro

Nakamura

Nakamura²,

Fukunaga³

2014

Cells Tissues Organs

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Perlecan is a multifunctional component of the extracellular matrix. It shows different effects on distinct cell types, and therefore it is thought to show potential for therapies targeting multiple cell types. However, the full range of multifunctionality of perlecan remains to be elucidated. We cultured various cell types, which were derived from epithelial/endothelial, connective and muscle tissues, in the presence of either antiserum against perlecan or exogenous perlecan, and examined the effects of perlecan on cell migration and proliferation. Cell migration was determined using a scratch assay. Blocking of perlecan by anti-perlecan antiserum inhibited the migration of vascular endothelial cells (VECs) and bone marrow-derived mesenchymal stem cells, and exogenous perlecan added to the culture medium promoted the migration of these cell types. The migration of other cell types was inhibited or was not promoted by exogenous perlecan. Cell proliferation was measured using a water-soluble tetrazolium dye. When cells were cultured at low densities, perlecan blocking inhibited the proliferation of VECs, and exogenous perlecan promoted the proliferation of keratinocytes. In contrast, the proliferation of fibroblasts, pre-adipocytes and vascular smooth muscle cells cultured at low densities was inhibited by exogenous perlecan. When cells were cultured at high densities, perlecan blocking promoted the proliferation of most cell types, with the exception of skeletal system-derived cells (chondrocytes and osteoblasts), which were inhibited by exogenous perlecan. Our results provide an overview of the multiple functions of perlecan in various cell types, and implicate a potential role of perlecan to inhibit undesirable activities, such as fibrosis, obesity and intimal hyperplasia.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Heparitinase-I could not be detected by CBB staining. The data are adapted from Nakamura et al [2014a].…”

Section: Methodsmentioning

confidence: 99%

Section: Dot Blot Analysismentioning

confidence: 99%

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Perlecan Diversely Regulates the Migration and Proliferation of Distinct Cell Types in vitro

Nakamura

Nakamura²,

Fukunaga³

2014

Cells Tissues Organs

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Heparan sulfate proteoglycans as key regulators of the mesenchymal niche of hematopoietic stem cells

Papy-García

Albanese

2017

Glycoconj J

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The complex microenvironment that surrounds hematopoietic stem cells (HSCs) in the bone marrow niche involves different coordinated signaling pathways. The stem cells establish permanent interactions with distinct cell types such as mesenchymal stromal cells, osteoblasts, osteoclasts or endothelial cells and with secreted regulators such as growth factors, cytokines, chemokines and their receptors. These interactions are mediated through adhesion to extracellular matrix compounds also. All these signaling pathways are important for stem cell fates such as self-renewal, proliferation or differentiation, homing and mobilization, as well as for remodeling of the niche. Among these complex molecular cues, this review focuses on heparan sulfate (HS) structures and functions and on the role of enzymes involved in their biosynthesis and turnover. HS associated to core protein, constitute the superfamily of heparan sulfate proteoglycans (HSPGs) present on the cell surface and in the extracellular matrix of all tissues. The key regulatory effects of major medullar HSPGs are described, focusing on their roles in the interactions between hematopoietic stem cells and their endosteal niche, and on their ability to interact with Heparin Binding Proteins (HBPs). Finally, according to the relevance of HS moieties effects on this complex medullar niche, we describe recent data that identify HS mimetics or sulfated HS signatures as new glycanic tools and targets, respectively, for hematopoietic and mesenchymal stem cell based therapeutic applications.

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Integrins and other cell surface attachment molecules of bone cells

Marie

Teti

2020

Principles of Bone Biology

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Contrasting effect of perlecan on adipogenic and osteogenic differentiation of mesenchymal stem cells in vitro

Cited by 14 publications

References 29 publications

Perlecan Diversely Regulates the Migration and Proliferation of Distinct Cell Types in vitro

Perlecan Diversely Regulates the Migration and Proliferation of Distinct Cell Types in vitro

Heparan sulfate proteoglycans as key regulators of the mesenchymal niche of hematopoietic stem cells

Integrins and other cell surface attachment molecules of bone cells

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