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Polymeric nanofibrous substrates stimulate pluripotent stem cells to form three-dimensional multilayered patty-like spheroids in feeder-free culture and maintain their pluripotency

Alamein, Mohammad A.; Wolvetang, Ernst J.; Ovchinnikov, Dmitry A.; Stephens, Sebastien; Sanders, Katherine; Warnke, Patrick H.

Authors

Mohammad A. Alamein

Ernst J. Wolvetang

Dmitry A. Ovchinnikov

Sebastien Stephens

Katherine Sanders

Patrick H. Warnke



Abstract

Expansion of pluripotent stem cells in defined media devoid of animal-derived feeder cells to generate multilayered three-dimensional (3D) bulk preparations or spheroids, rather than two-dimensional (2D) monolayers, is advantageous for many regenerative, biological or disease-modelling studies. Here we show that electrospun polymer matrices comprised of nanofibres that mimic the architecture of the natural fibrous extracellular matrix allow for feeder-free expansion of pluripotent human induced pluripotent stem cells (IPSCs) and human embryonic stem cells (HESCs) into multilayered 3D 'patty-like' spheroid structures in defined xeno-free culture medium. The observation that IPSCs and HESCs readily revert to 2D growth in the absence of the synthetic nanofibre membranes suggests that this 3D expansion behaviour is mediated by the physical microenvironment and artificial niche provided by the nanofibres only. Importantly, we could show that such 3D growth as patties maintained the pluripotency of cells as long as they were kept on nanofibres. The generation of complex multilayered 3D structures consisting of only pluripotent cells on biodegradable nanofibre matrices of the desired shape and size will enable both industrial-scale expansion and intricate organ-tissue engineering applications with human pluripotent stem cells, where simultaneous coupling of differentiation pathways of all germ layers from one stem cell source may be required for organ formation.

Citation

Alamein, M. A., Wolvetang, E. J., Ovchinnikov, D. A., Stephens, S., Sanders, K., & Warnke, P. H. (2015). Polymeric nanofibrous substrates stimulate pluripotent stem cells to form three-dimensional multilayered patty-like spheroids in feeder-free culture and maintain their pluripotency. Journal of tissue engineering and regenerative medicine, 9(9), 1078-1083. https://doi.org/10.1002/term.1960

Journal Article Type Article
Acceptance Date Sep 17, 2014
Online Publication Date Nov 25, 2014
Publication Date Sep 1, 2015
Deposit Date Aug 23, 2023
Journal Journal of Tissue Engineering and Regenerative Medicine
Print ISSN 1932-6254
Electronic ISSN 1932-7005
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 9
Issue 9
Pages 1078-1083
DOI https://doi.org/10.1002/term.1960
Keywords Nanofibres; Electrospinning; PMEDSAH; PLGA; Induced pluripotent stem cells (IPSCs); Human embryonic stem cells (HESCs); Pluripotency; Three-dimensional; Feeder-free; Xeno-free; Extracellular matrix
Public URL https://hull-repository.worktribe.com/output/4366654