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Directing human embryonic stem cell differentiation by non-viral delivery of siRNA in 3D culture

Author(s)
Lytton-Jean, Abigail K. R.; Deiorio-Haggar, Kaila; Bellan, Leon M.; Karagiannis, Emmanouil; Anderson, Daniel Griffith; Zoldan, Janeta; Langer, Robert S; ... Show more Show less
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Abstract
Human embryonic stem cells (hESCs) hold great potential as a resource for regenerative medicine. Before achieving therapeutic relevancy, methods must be developed to control stem cell differentiation. It is clear that stem cells can respond to genetic signals, such as those imparted by nucleic acids, to promote lineage-specific differentiation. Here we have developed an efficient system for delivering siRNA to hESCs in a 3D culture matrix using lipid-like materials. We show that non-viral siRNA delivery in a 3D scaffolds can efficiently knockdown 90% of GFP expression in GFP-hESCs. We further show that this system can be used as a platform for directing hESC differentiation. Through siRNA silencing of the KDR receptor gene, we achieve concurrent downregulation (60–90%) in genes representative of the endoderm germ layer and significant upregulation of genes representative of the mesoderm germ layer (27–90 fold). This demonstrates that siRNA can direct stem cell differentiation by blocking genes representative of one germ layer and also provides a particularly powerful means to isolate the endoderm germ layer from the mesoderm and ectoderm. This ability to inhibit endoderm germ layer differentiation could allow for improved control over hESC differentiation to desired cell types.
Date issued
2011-08
URI
http://hdl.handle.net/1721.1/99182
Department
Harvard University--MIT Division of Health Sciences and Technology; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Chemical Engineering; Massachusetts Institute of Technology. Media Laboratory; Koch Institute for Integrative Cancer Research at MIT
Journal
Biomaterials
Publisher
Elsevier
Citation
Zoldan, Janet, Abigail K.R. Lytton-Jean, Emmanouil D. Karagiannis, Kaila Deiorio-Haggar, Leon M. Bellan, Robert Langer, and Daniel G. Anderson. “Directing Human Embryonic Stem Cell Differentiation by Non-Viral Delivery of siRNA in 3D Culture.” Biomaterials 32, no. 31 (November 2011): 7793–7800.
Version: Author's final manuscript
ISSN
01429612
1878-5905

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