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Stem Cell Reviews and Reports

, Volume 12, Issue 4, pp 476–483 | Cite as

Functionalizing Ascl1 with Novel Intracellular Protein Delivery Technology for Promoting Neuronal Differentiation of Human Induced Pluripotent Stem Cells

  • Meghan Robinson
  • Parv Chapani
  • Tara Styan
  • Ranjani Vaidyanathan
  • Stephanie Michelle WillerthEmail author
Article

Abstract

Pluripotent stem cells can become any cell type found in the body. Accordingly, one of the major challenges when working with pluripotent stem cells is producing a highly homogenous population of differentiated cells, which can then be used for downstream applications such as cell therapies or drug screening. The transcription factor Ascl1 plays a key role in neural development and previous work has shown that Ascl1 overexpression using viral vectors can reprogram fibroblasts directly into neurons. Here we report on how a recombinant version of the Ascl1 protein functionalized with intracellular protein delivery technology (Ascl1-IPTD) can be used to rapidly differentiate human induced pluripotent stem cells (hiPSCs) into neurons. We first evaluated a range of Ascl1-IPTD concentrations to determine the most effective amount for generating neurons from hiPSCs cultured in serum free media. Next, we looked at the frequency of Ascl1-IPTD supplementation in the media on differentiation and found that one time supplementation is sufficient enough to trigger the neural differentiation process. Ascl1-IPTD was efficiently taken up by the hiPSCs and enabled rapid differentiation into TUJ1-positive and NeuN-positive populations with neuronal morphology after 8 days. After 12 days of culture, hiPSC-derived neurons produced by Ascl1-IPTD treatment exhibited greater neurite length and higher numbers of branch points compared to neurons derived using a standard neural progenitor differentiation protocol. This work validates Ascl1-IPTD as a powerful tool for engineering neural tissue from pluripotent stem cells.

Keywords

Stem cells Proteins Drug delivery Neuroscience Intracellular targeting Neurite extension 

Notes

Acknowledgments

The authors would like to acknowledge support from our NSERC Engage grant with iProgen Biotech, Inc. and funding from the Canada Research Chairs Program (S.M.W.).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Meghan Robinson
    • 1
  • Parv Chapani
    • 2
  • Tara Styan
    • 1
  • Ranjani Vaidyanathan
    • 3
  • Stephanie Michelle Willerth
    • 4
    • 5
    • 6
    Email author
  1. 1.Biomedical Engineering ProgramUniversity of VictoriaVictoriaCanada
  2. 2.Department of BiochemistryUniversity of VictoriaVictoriaCanada
  3. 3.iProgen Biotech, Inc.VancouverCanada
  4. 4.Department of Mechanical EngineeringUniversity of VictoriaVictoriaCanada
  5. 5.Division of Medical SciencesUniversity of VictoriaVictoriaCanada
  6. 6.International Collaboration on Repair Discoveries (ICORD)VancouverCanada

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