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Cited by 47RNA Interference and Gene DeliveryPluripotent Stem Cells ResearchMesenchymal stem cell research

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

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Zoldan, Janet, Lytton-Jean, Abigail K.R., Karagiannis, Emmanouil D., Deiorio-Haggar, Kaila, Bellan, Leon M., Langer, Robert, Anderson, Daniel G.·2011·Biomaterials
DOI 10.1016/j.biomaterials.2011.06.057PMC PMC3313656
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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.

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  • Gene-activated matrices maintain permanent transfection with 60-90% siRNA-mediated gene knock-down during 3D organoid development but do not allow transfection at different time points.

    “While this system maintains a permanent transfection with 60–90% of siRNA-mediated gene knock-down during 3D organoid development, it does not allow transfection at different time points.”

    From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [18]

    The Introduction reports 60–90% silencing in a 3D hESC matrix, and the Results report siRNA-mediated knockdown during differentiation. But the Results describe knockdown measured after three days, not permanent transfection or 3D organoid development. The paper does not establish that transfection cannot occur at different time points. (LLM, conf=0.88)

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Research works printed but not linked: 43

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Authors
Zoldan, J., Lytton-Jean, A. K. R., Karagiannis, E. D., Deiorio-Haggar, K., Bellan, L. M., Langer, R., & Anderson, D. G.
Affiliations
  • Koch Institute for Integrative Cancer Research At MIT (US)
  • Massachusetts Institute of Technology (US)
  • Harvard–MIT Division of Health Sciences and Technology (US)
Corresponding author
Anderson, Daniel G.
ORCID
Venue
Biomaterials
Publisher
Elsevier BV
Published
2011-11
Volume
32
Issue
31
Pages
7793-7800
ISSN
0142-9612 (ISSN-L)
DOI
10.1016/j.biomaterials.2011.06.057
PMC
PMC3313656
OpenAlex
W1996740541
Cited by
47
References
0
Citations by year
2026: 1 · 2025: 1 · 2023: 1 · 2021: 2 · 2020: 3 · 2019: 4 · 2018: 3 · 2017: 1 · 2016: 6 · 2015: 5 · 2014: 6 · 2013: 8 · 2012: 5
FWCI
2.60
Type
journal article
Language
en
Open access
Licence
Locations
  • Biomaterials (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-nd)
Funders
  • Alnylam Pharmaceuticals
  • National Institutes of Health (5r01de016516-10, 4r37eb000244-34, EB-000244)
  • National Institute of Biomedical Imaging and Bioengineering (EB-000244, R01EB000244, R37-EB000244)
  • National Institute of Dental and Craniofacial Research (R01-DE016516, DE016561)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
Keywords
human embryonic stem cells, siRNA delivery, non-viral gene delivery, 3D cell culture, stem cell differentiation, germ layer specification, endoderm differentiation, mesoderm differentiation, KDR receptor, gene silencing, regenerative medicine
MeSH terms
Animals, Cell Differentiation, Cells, Cultured, Humans, Viruses, Gene Transfer Techniques, Cell Culture Techniques, Reverse Transcriptase Polymerase Chain Reaction, RNA, Small Interfering, Green Fluorescent Proteins, Mice, Embryonic Stem Cells, Gene Knockdown Techniques, Embryoid Bodies, Protein Interaction Maps

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