Fibronectin-mediated upregulation of α5β1 integrin and cell adhesion during differentiation of mouse embryonic stem cells

Pimton, Pimchanok; Sarkar, Saheli; Sheth, Nidhi; Perets, Anat; Marcinkiewicz, Cezary; Lazarovici, Philip; Lelkes, Peter I.

doi:10.4161/cam.5.1.13704

Cited by 38 publications

(31 citation statements)

References 40 publications

(42 reference statements)

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“…This result was initially confusing given the large body of data implicating VEGF signaling in mesoderm and EC fate [7, 8, 37, 58, 59], however, the results provided in this manuscript examining 4 ESC lines indicate that BMP-4 signaling on fibronectin are adequate signals for population-based EPC and EC inductions. We expect that the two distinct VEGF binding domains within FN matrix [48] is sequestering the autocrine VEGF produced by the cells (Fig 8D) and may even be protecting the VEGF from degradation [49, 50]. Our results advocate that the utilization of FN matrix is superior to collagen IV and usually mitigates the need for VEGF supplementation in both VPC and EC induction cultures.…”

Section: Discussionmentioning

confidence: 89%

“…The extracellular matrix (ECM) protein, fibronectin (FN), has been shown to contain two novel VEGF binding domains using direct physical association between the VEGF receptor-2 (Flk-1) and the FN integrin, alpha5beta1 [48] to activate cell signaling cascades FN, thus directing EC fate by sequestering VEGF in an insoluble form [49, 50]. When mixed directly with the FN and incubated overnight, VEGF can also be visualized from day 0–3 (not shown) and an ELISA assay found no detectable VEGF released into the aspirate during the course of the 3-day period (not shown).…”

Section: Resultsmentioning

confidence: 99%

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Multifactorial Optimizations for Directing Endothelial Fate from Stem Cells

et al. 2016

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Embryonic stem cells (ESC) and induced pluripotent stem (iPS) cells are attractive in vitro models of vascular development, therapeutic angiogenesis, and tissue engineering. However, distinct ESC and iPS cell lines respond differentially to the same microenvironmental factors. Developing improved/optimized differentiation methodologies tailored/applicable in a number of distinct iPS and ESC lines remains a challenge in the field. Currently published methods for deriving endothelial cells (EC) robustly generate high numbers of endothlelial progenitor cells (EPC) within a week, but their maturation to definitive EC is much more difficult, taking up to 2 months and requiring additional purification. Therefore, we set out to examine combinations/levels of putative EC induction factors—utilizing our stage-specific chemically-defined derivation methodology in 4 ESC lines including: kinetics, cell seeding density, matrix signaling, as well as medium treatment with vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF). The results indicate that temporal development in both early and late stages is the most significant factor generating the desired cells. The generation of early Flk-1+/KDR+ vascular progenitor cells (VPC) from pluripotent ESC is directed predominantly by high cell seeding density and matrix signaling from fibronectin, while VEGF supplementation was NOT statistically significant in more than one cell line, especially with fibronectin matrix which sequesters autocrine VEGF production by the differentiating stem cells. Although some groups have shown that the GSK3-kinase inhibitor (CHIR) can facilitate EPC fate, it hindered the generation of KDR+ cells in our preoptimized medium formulations. The methods summarized here significantly increased the production of mature vascular endothelial (VE)-cadherin+ EC, with up to 93% and 57% purity from mouse and human ESC, respectively, before VE-cadherin+ EC purification.

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Section: Discussionmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Multifactorial Optimizations for Directing Endothelial Fate from Stem Cells

et al. 2016

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“…Laminin-322 stimulates osteogenic differentiation of mesenchymal stem cells, whereas laminin-111 promotes neural differentiation (Klees et al, 2005;Klees et al, 2007;Mruthyunjaya et al, 2010). Fibronectin can mediate ES cell differentiation towards a meso-endodermal lineage (Hayashi et al, 2007) by upregulating the expression of integrin a5b1 (Pimton et al, 2011). Under different culture conditions, however, fibronectin promotes ES cell self-renewal (Hunt et al, 2012).…”

Section: The Ecm In Stem Cell Differentiationmentioning

confidence: 99%

Fibronectin and stem cell differentiation – lessons from chondrogenesis

Singh

Schwarzbauer

2012

Journal of Cell Science

168

156

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SummaryThe extracellular matrix (ECM) is an intricate network of proteins that surrounds cells and has a central role in establishing an environment that is conducive to tissue-specific cell functions. In the case of stem cells, this environment is the stem cell niche, where ECM signals participate in cell fate decisions. In this Commentary, we describe how changes in ECM composition and mechanical properties can affect cell shape and stem cell differentiation. Using chondrogenic differentiation as a model, we examine the changes in the ECM that occur before and during mesenchymal stem cell differentiation. In particular, we focus on the main ECM protein fibronectin, its temporal expression pattern during chondrogenic differentiation, its potential effects on functions of differentiating chondrocytes, and how its interactions with other ECM components might affect cartilage development. Finally, we discuss data that support the possibility that the fibronectin matrix has an instructive role in directing cells through the condensation, proliferation and/or differentiation stages of cartilage formation.

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“…Furthermore, it is generally agreed that the immunodepletion of the hematopoietic cells using anti-CD11b, CD34, and CD45, forms the fundamental concept for negative selection method of MSCs purification (26,27). In addition, the systemic analysis of cell surface molecules of MSCs has revealed that MSCs also express receptors for numerous extracellular matrix (ECM) proteins including collagen (α1β1-, α2β1-integrin), laminin (α6β1-, α6β4-integrin), fibronectin (α3β1-, α5β1-integrin), and vitronectin (αvβ1-, αvβ3-integrin) (28)(29)(30)(31). These specific expression patterns of adhesion molecules suggest the potential interactions between MSCs and other cell types in vivo (32)(33)(34).…”

Section: Characterizationmentioning

confidence: 99%

Mesenchymal Stem Cells and Nano-structured Surfaces

Zhou

Chakravorty

Xiao

et al. 2013

Methods in Molecular Biology

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Mesenchymal stem cells (MSCs) represent multipotent stromal cells that can differentiate into a variety of cell types, including osteoblasts (bone cells), chondrocytes (cartilage cells), and adipocytes (fat cells). Their multi-potency provides a great promise as a cell source for tissue engineering and cell-based therapy for many diseases, particularly bone diseases and bone formation. To be able to direct and modulate the differentiation of MSCs into the desired cell types in situ in the tissue, nanotechnology is introduced and used to facilitate or promote cell growth and differentiation. These nano-materials can provide a fine structure and tuneable surface in nanoscales to help the cell adhesion and promote the cell growth and differentiation of MSCs. This could be a dominant direction in future for stem cells based therapy or tissue engineering for various diseases. Therefore, the isolation, manipulation, and differentiation of MSCs are very important steps to make meaningful use of MSCs for disease treatments. In this chapter, we have described a method of isolating MSC from human bone marrow, and how to culture and differentiate them in vitro. We have also provided research methods on how to use MSCs in an in vitro model and how to observe MSC biological response on the surface of nano-scaled materials.

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Fibronectin-mediated upregulation of α5β1 integrin and cell adhesion during differentiation of mouse embryonic stem cells

Cited by 38 publications

References 40 publications

Multifactorial Optimizations for Directing Endothelial Fate from Stem Cells

Multifactorial Optimizations for Directing Endothelial Fate from Stem Cells

Fibronectin and stem cell differentiation – lessons from chondrogenesis

Mesenchymal Stem Cells and Nano-structured Surfaces

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