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

Jessie L. Ratcliffe, Marc Walker, Ahmed M. Eissa, Shengrong Du, Stefan A. Przyborski, Andrew L. Laslett, Neil R. Cameron

Research output: Contribution to journalArticleResearchpeer-review

2 Citations (Scopus)

Abstract

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.

Original languageEnglish
Number of pages8
JournalJournal of Polymer Science, Part A: Polymer Chemistry
DOIs
Publication statusAccepted/In press - 9 Mar 2019

Keywords

  • 3D cell culture
  • emulsions
  • photopolymerization
  • porous polymers
  • surface functionalization

Cite this

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title = "Optimized peptide functionalization of thiol-acrylate emulsion-templated porous polymers leads to expansion of human pluripotent stem cells in 3D culture",
abstract = "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.",
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author = "Ratcliffe, {Jessie L.} and Marc Walker and Eissa, {Ahmed M.} and Shengrong Du and Przyborski, {Stefan A.} and Laslett, {Andrew L.} and Cameron, {Neil R.}",
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Optimized peptide functionalization of thiol-acrylate emulsion-templated porous polymers leads to expansion of human pluripotent stem cells in 3D culture. / Ratcliffe, Jessie L.; Walker, Marc; Eissa, Ahmed M.; Du, Shengrong; Przyborski, Stefan A.; Laslett, Andrew L.; Cameron, Neil R.

In: Journal of Polymer Science, Part A: Polymer Chemistry, 09.03.2019.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Optimized peptide functionalization of thiol-acrylate emulsion-templated porous polymers leads to expansion of human pluripotent stem cells in 3D culture

AU - Ratcliffe, Jessie L.

AU - Walker, Marc

AU - Eissa, Ahmed M.

AU - Du, Shengrong

AU - Przyborski, Stefan A.

AU - Laslett, Andrew L.

AU - Cameron, Neil R.

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