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Article

Self-Assembling Scaffolds Supported Long-Term Growth of Human Primed Embryonic Stem Cells and Upregulated Core and Naïve Pluripotent Markers

1
Department of Biological Sciences, Oakland University, Rochester, MI 48309, USA
2
OU-WB Institute for Stem Cell and Regenerative Medicine, Rochester, MI 48309, USA
*
Author to whom correspondence should be addressed.
Cells 2019, 8(12), 1650; https://doi.org/10.3390/cells8121650
Received: 5 October 2019 / Revised: 13 December 2019 / Accepted: 14 December 2019 / Published: 16 December 2019
(This article belongs to the Special Issue 3D Stem Cell Culture)
The maintenance and expansion of human embryonic stem cells (ESCs) in two-dimensional (2-D) culture is technically challenging, requiring routine manipulation and passaging. We developed three-dimensional (3-D) scaffolds to mimic the in vivo microenvironment for stem cell proliferation. The scaffolds were made of two 8-arm polyethylene glycol (PEG) polymers functionalized with thiol (PEG-8-SH) and acrylate (PEG-8-Acr) end groups, which self-assembled via a Michael addition reaction. When primed ESCs (H9 cells) were mixed with PEG polymers, they were encapsulated and grew for an extended period, while maintaining their viability, self-renewal, and differentiation potential both in vitro and in vivo. Three-dimensional (3-D) self-assembling scaffold-grown cells displayed an upregulation of core pluripotency genes, OCT4, NANOG, and SOX2. In addition, the expression of primed markers decreased, while the expression of naïve markers substantially increased. Interestingly, the expression of mechanosensitive genes, YAP and TAZ, was also upregulated. YAP inhibition by Verteporfin abrogated the increased expression of YAP/TAZ as well as core and naïve pluripotent markers. Evidently, the 3-D culture conditions induced the upregulation of makers associated with a naïve state of pluripotency in the primed cells. Overall, our 3-D culture system supported the expansion of a homogenous population of ESCs and should be helpful in advancing their use for cell therapy and regenerative medicine. View Full-Text
Keywords: embryonic stem cells; three-dimensional; self-assembling scaffold; pluripotency; culture conditions; expansion; growth; niche embryonic stem cells; three-dimensional; self-assembling scaffold; pluripotency; culture conditions; expansion; growth; niche
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MDPI and ACS Style

McKee, C.; Brown, C.; Chaudhry, G.R. Self-Assembling Scaffolds Supported Long-Term Growth of Human Primed Embryonic Stem Cells and Upregulated Core and Naïve Pluripotent Markers. Cells 2019, 8, 1650. https://doi.org/10.3390/cells8121650

AMA Style

McKee C, Brown C, Chaudhry GR. Self-Assembling Scaffolds Supported Long-Term Growth of Human Primed Embryonic Stem Cells and Upregulated Core and Naïve Pluripotent Markers. Cells. 2019; 8(12):1650. https://doi.org/10.3390/cells8121650

Chicago/Turabian Style

McKee, Christina, Christina Brown, and G. Rasul Chaudhry. 2019. "Self-Assembling Scaffolds Supported Long-Term Growth of Human Primed Embryonic Stem Cells and Upregulated Core and Naïve Pluripotent Markers" Cells 8, no. 12: 1650. https://doi.org/10.3390/cells8121650

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