Long-Term Maintenance of Human Pluripotent Stem Cells on cRGDfK-Presenting Synthetic Surfaces

Lambshead, Jack; Meagher, Laurence; Goodwin, Jacob; Labonne, Tanya; Ng, Elizabeth; Elefanty, Andrew G.; O'Brien, Carmel; Laslett, Andrew L.

doi:10.1038/s41598-018-19209-0

Cited by 27 publications

(22 citation statements)

References 59 publications

(72 reference statements)

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“…The synthetic ligand interacted with human induced pluripotent stem cells (hIPSCs) via the integrin a v b 5 units, demonstrating its comparability to vitronectin. Lambshead et al observed human pluripotent stem cells (hPSCs) cultured on Synthemax ® coated plates and flasks were morphologically indistinguishable from those cultured in control flasks coated with Geltrex (Lambshead et al, 2018). Accordingly, the genetic stability and pluripotency of hPSCs was maintained on Synthemax ® surface as assessed by the PluriTest ™ assay (Muller et al, 2011).…”

Section: Synthetic Peptide Surface Microcarrier: Synthemax ®mentioning

confidence: 99%

Biological Considerations in Scaling Up Therapeutic Cell Manufacturing

et al. 2020

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Cell therapeuticsusing cells as living drugshave made advances in many areas of medicine. One of the most clinically studied cell-based therapy products is mesenchymal stromal cells (MSCs), which have shown promising results in promoting tissue regeneration and modulating inflammation. However, MSC therapy requires large numbers of cells, the generation of which is not feasible via conventional planar tissue culture methods. Scale-up manufacturing methods (e.g., propagation on microcarriers in stirred-tank bioreactors), however, are not specifically tailored for MSC expansion. These processes may, in principle, alter the cell secretome, a vital component underlying the immunosuppressive properties and clinical effectiveness of MSCs. This review outlines our current understanding of MSC properties and immunomodulatory function, expansion in commercial manufacturing systems, and gaps in our knowledge that need to be addressed for effective up-scaling commercialization of MSC therapy.

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Section: Synthetic Peptide Surface Microcarrier: Synthemax ®mentioning

confidence: 99%

Biological Considerations in Scaling Up Therapeutic Cell Manufacturing

et al. 2020

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“…42 In this paper, we explore the functionalization of thiol-acrylate polyHIPE materials with the peptide cyclo-arginine-glycineaspartate (c-RGD), which has been shown to lead to the expansion of human pluripotent stem cells in 2D culture. 43 In this manner, we aim to produce enhanced materials for in vitro pluripotent stem cell culture and expansion in 3D. We utilize the residual thiols in the polyHIPE materials to react with maleimidefunctionalized c-RGD-mal, via a Michael addition.…”

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confidence: 99%

Optimized peptide functionalization of thiol‐acrylate emulsion‐templated porous polymers leads to expansion of human pluripotent stem cells in 3D culture

Ratcliffe

Walker

Eissa

et al. 2019

J. Polym. Sci. Part A: Polym. Chem.

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Highly porous polymers produced by polymerization of the continuous phase of a high internal phase emulsion have been developed as scaffolds for 3D culture of human pluripotent stem cells. These emulsion‐templated polymerized high internal phase emulsion (polyHIPE) materials have an interconnected network of pores that provide support for the cells, while also allowing both cell ingress and nutrient diffusion. Thiol‐acrylate polyHIPE materials were prepared by photopolymerization, which, due to a competing acrylate homopolymerization process, leads to a material with residual surface thiols. These thiols were then used as a handle to allow postpolymerization functionalization with both maleimide and a maleimide‐derivatized cyclo‐RGDfK peptide, via Michael addition under benign conditions. Functionalization was evaluated using an Ellman's colorimetric assay, to monitor the residual thiol concentration, and X‐ray photoelectron spectroscopy. Maleimide was used as a model molecule to optimize conditions prior to peptide‐functionalization. The use of triethylamine as a catalyst and a mixed ethanol‐aqueous solvent system led to optimized reaction between surface‐bound thiols and maleimide. Peptide‐functionalized materials showed improved attachment and infiltration of human pluripotent stem cells over 7 days, demonstrating their promise as a scaffold for 3D stem cell culture and expansion. © 2019 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2019, 57, 1974–1981

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“…Integrin-mediated hPSC attachment to synthetic polymer substrates, notably of β1 and αv integrins, has been previously reported. [11,23] These data suggest that αvβ3 and αvβ5 heterodimers play an important role for mediating hPSC attachment to poly (TCDMDA-blend-BA), however, integrins are likely to interact in a complex manner and therefore can form a number of different homo-or hetero-dimers.…”

Section: Main Sectionmentioning

confidence: 96%

Discovery of a Novel Polymer for Xeno-free, Long-term Culture of Human Pluripotent Stem Cell Expansion

Thorpe

Nasir

Burroughs³

et al. 2020

Preprint

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Human pluripotent stem cells (hPSCs) can be expanded and differentiated in vitro into almost any adult tissue cell type, and thus have great potential as a source for cell therapies with biomedical application. In this study, a fully-defined polymer synthetic substrate is identified for hPSC culture in completely defined, xeno-free conditions. This system can overcome the cost, scalability and reproducibility limitations of current hPSC culture strategies, and facilitate large-scale production. A high-throughput, multi-generational polymer microarray platform approach was used to test over 600 unique polymers and rapidly assess hPSC-polymer interactions in combination with the fully defined xeno-free medium, Essential 8TM (E8). This study identifies as novel nanoscale phase separated blend of poly(tricyclodecane-dimethanol diacrylate) and poly(butyl acrylate) (2:1 v/v), which supports long-term expansion of hPSCs and can be readily coated onto standard cultureware. Analysis of cell-polymer interface interactions through mass spectrometry and integrin blocking studies provides novel mechanistic insight into the role of the E8 proteins in promoting integrin-mediated hPSC attachment and maintaining hPSC signaling, including ability to undergo multi-lineage differentiation. This study therefore identifies a novel substrate for long-term serial passaging of hPSCs in serum-free, commercial chemically-defined E8, which provides a promising and economic hPSC expansion platform for clinical-scale application.

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Long-Term Maintenance of Human Pluripotent Stem Cells on cRGDfK-Presenting Synthetic Surfaces

Cited by 27 publications

References 59 publications

Biological Considerations in Scaling Up Therapeutic Cell Manufacturing

Biological Considerations in Scaling Up Therapeutic Cell Manufacturing

Optimized peptide functionalization of thiol‐acrylate emulsion‐templated porous polymers leads to expansion of human pluripotent stem cells in 3D culture

Discovery of a Novel Polymer for Xeno-free, Long-term Culture of Human Pluripotent Stem Cell Expansion

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