Directing human embryonic stem cell differentiation by non-viral delivery of siRNA in 3D culture
DOI: 10.1016/j.biomaterials.2011.06.057
Title: Directing human embryonic stem cell differentiation by non-viral delivery of siRNA in 3D culture
Journal Title: Biomaterials
Volume: 32
Issue: 31
Publication Date: November 2011
Start Page: 7793
End Page: 7800
Published online: online 11 August 2011
ISSN: 0142-9612
Affiliations:

  • a David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA

  • b Department of Biology, Massachusetts Institute of Technology, Cambridge MA, USA

  • c Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge MA, USA

  • d Harvard-MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA
  • Abstract: bryonic 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.
    Accepted: 24 June 2011
    Received: 28 April 2011
    Tel: +617 258 6843
    Email: dgander@mit.edu

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