Synthetic polymer coatings for long-term growth of human embryonic stem cells

Villa‐Diaz, Luis G.; Nandivada, Himabindu; Ding, Jun; Nogueira-de-Souza, Naiara Correa; Krebsbach, Paul H.; O’Shea, K. Sue; Lahann, Joerg; Smith, Gary D.

doi:10.1038/nbt.1631

Cited by 332 publications

(266 citation statements)

References 12 publications

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“…Another commercially available culture media StemPro was used to maintain HUES9 and HUES1 cells for five passages on the synthetic polymer poly(methylvinylether-alt-maleic anhydride) [30]. The use of synthetic polymers with variable wettability and rigidity has been evaluated to support hES cell expansion in conditioned medium in short-term cultures but long-term propagations on these surfaces failed in defined, serum-free medium [31]. The use of ECM components or undefined culture media, for example conditioned media, makes it difficult to determine how the defined ECM components alone are influencing the hES cell response.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the interface between adsorbed fibronectin and human embryonic stem cells

Kalaskar

Downes

Murray

et al. 2013

J. R. Soc. Interface.

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The cell-substrate interface plays a key role in the regulation of cell behaviour. Defining the properties of this interface is particularly important for human embryonic stem (hES) cell culture, because changes in this environment can regulate hES cell differentiation. It has been established that fibronectincoated surfaces can promote the attachment, growth and maintenance of the undifferentiated phenotype of hES cells. We investigated the influence of the surface density of adsorbed fibronectin on hES cell behaviour in defined serum-free culture conditions and demonstrated that only 25 per cent surface saturation was required to maintain attachment, growth and maintenance of the undifferentiated phenotype. The influence of surface-adsorbed fibronectin fragments was compared with whole fibronectin, and it was demonstrated that the 120 kDa fragment central binding domain alone was able to sustain hES cells in an undifferentiated phenotype in a similar fashion to fibronectin. Furthermore, hES cell attachment to both fibronectin and the 120 kDa fragment was mediated by integrin a5b1. However, although a substrateattached synthetic arginine-glycine-aspartic acid (RGD) peptide alone was able to promote the attachment and spreading of fibroblasts, it was inactive for hES cells, indicating that stem cells have different requirements in order to attach and spread on the central fibronectin RGD-cell-binding domain. This study provides further information on the characteristics of the cellsubstrate interface required to control hES cell behaviour in clearly defined serum-free conditions, which are needed for the development of therapeutic applications of hES cells.

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Section: Discussionmentioning

confidence: 99%

Characterization of the interface between adsorbed fibronectin and human embryonic stem cells

Kalaskar

Downes

Murray

et al. 2013

J. R. Soc. Interface.

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“…Here, we develop a technology that notably provides expansion rates greater than the gold standard of feeder substrates; works with enzymatic passaging; and is compatible with current manipulations involving hESCs/hiPSCs, including gene modification and reprogramming. Importantly, in contrast to the previous reports that used polymers and/or peptides to coat the dish (10)(11)(12)14), our UV/ozone (UV) treatment of common research materials is an attractive alternative because it is inexpensive and mass production of appropriately treated culture dishes for numerous applications should be straightforward.…”

mentioning

confidence: 92%

“…A variety of substrates are used in these methods: synthetic polymers (11,14), peptide-modified surfaces (10,12), embryonic ECM laminin isoforms (7), fibronectin from ECM with a small molecule mixture (9), and various vitronectin proteins (4,6). Some of these culture methods use animal products, which makes potential transplantation applications problematic (21).…”

mentioning

confidence: 99%

“…induced pluripotent stem cells (hiPSCs) have the potential to grow indefinitely in culture and to make any cell in the adult human body, they represent important resources for regenerative medicine (1) and for disease modeling efforts that seek to recreate a patient's disease pathogenesis in the laboratory (2,3). hESCs and hiPSCs are typically grown on a "feeder" cell layer of mitotically inactivated mouse embryonic fibroblasts (mEFs) or on "feeder-free" culture systems composed of a variety of ECM/ serum proteins coated onto tissue culture dishes (4)(5)(6)(7)(8)(9) or synthetic materials (10)(11)(12)(13)(14). Nearly all these systems have been reported to promote hESC/hiPSC self-renewal when the cells are passaged in multicellular clumps and seeded at a suitably high cell density.…”

mentioning

confidence: 99%

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Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions

Saha

Mei²,

Reisterer³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

135

109

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The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.

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“…Such defined substrata have been developed for the long-term culture of undifferentiated hPS cells in defined media. A number of surfaces have been described including recombinant proteins (14)(15)(16), fully synthetic polymers (17)(18)(19), and peptide-modified surfaces (20)(21)(22)(23). Surfaces that present bioactive peptides have the advantage that they can be programmed to interact with specific cell-surface macromolecules such as the glycosaminoglycans (GAGs) and integrins.…”

mentioning

confidence: 99%

Signals from the surface modulate differentiation of human pluripotent stem cells through glycosaminoglycans and integrins

Wrighton¹,

Klim

Hernandez³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The fate decisions of human pluripotent stem (hPS) cells are governed by soluble and insoluble signals from the microenvironment. Many hPS cell differentiation protocols use Matrigel, a complex and undefined substrate that engages multiple adhesion and signaling receptors. Using defined surfaces programmed to engage specific cell-surface ligands (i.e., glycosaminoglycans and integrins), the contribution of specific matrix signals can be dissected. For ectoderm and motor neuron differentiation, peptide-modified surfaces that can engage both glycosaminoglycans and integrins are effective. In contrast, surfaces that interact selectively with glycosaminoglycans are superior to Matrigel in promoting hPS cell differentiation to definitive endoderm and mesoderm. The modular surfaces were used to elucidate the signaling pathways underlying these differences. Matrigel promotes integrin signaling, which in turn inhibits mesendoderm differentiation. The data indicate that integrin-activating surfaces stimulate Akt signaling via integrinlinked kinase (ILK), which is antagonistic to endoderm differentiation. The ability to attribute cellular responses to specific interactions between the cell and the substrate offers new opportunities for revealing and controlling the pathways governing cell fate.

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Synthetic polymer coatings for long-term growth of human embryonic stem cells

Cited by 332 publications

References 12 publications

Characterization of the interface between adsorbed fibronectin and human embryonic stem cells

Characterization of the interface between adsorbed fibronectin and human embryonic stem cells

Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions

Signals from the surface modulate differentiation of human pluripotent stem cells through glycosaminoglycans and integrins

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