Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Extracellular cues accelerate neurogenesis of induced pluripotent stem cell derived neurons

Elizabeth R. Sharlow, Danielle C. Llaneza, Anna J. Mendelson, Garnett A. Mingledorff, Veronica M. Porterfield, Kathryn Salvati, View ORCID ProfileMark Beenhakker, George S. Bloom, John S. Lazo
doi: https://doi.org/10.1101/2021.08.17.456634
Elizabeth R. Sharlow
1Department of Pharmacology, University of Virginia, Charlottesville, VA 22908
2Fiske Drug Discovery Laboratory, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: ers7g@virginia.edu
Danielle C. Llaneza
1Department of Pharmacology, University of Virginia, Charlottesville, VA 22908
2Fiske Drug Discovery Laboratory, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Anna J. Mendelson
1Department of Pharmacology, University of Virginia, Charlottesville, VA 22908
2Fiske Drug Discovery Laboratory, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Garnett A. Mingledorff
3University of Virginia Stem Cell Facility, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Veronica M. Porterfield
3University of Virginia Stem Cell Facility, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kathryn Salvati
1Department of Pharmacology, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mark Beenhakker
1Department of Pharmacology, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Mark Beenhakker
George S. Bloom
4Departments of Biology, Cell Biology and Neuroscience, University of Virginia, Charlottesville, VA 22904
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John S. Lazo
1Department of Pharmacology, University of Virginia, Charlottesville, VA 22908
2Fiske Drug Discovery Laboratory, University of Virginia, Charlottesville, VA 22908
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

Neurogenesis is a complex process encompassing neuronal progenitor cell expansion/proliferation and differentiation, followed by neuron maturation. In vivo models are most commonly used to study neurogenesis; however, human induced pluripotent stem cell-derived (iPSC) neurons are increasingly used to establish cellular models of human neurological processes. Unfortunately, the differentiation and maturation of iPSC-derived neurons varies in methodology, is asynchronous, and has restricted experimental utility because of extended differentiation/maturation times. To accelerate and standardize iPS neuronal maturation, we differentiated and matured feeder layer-free iPSC-derived neuronal cultures under physiological oxygen levels (5%), and modified the underlying extracellular matrix and medium composition. Our results demonstrate that calretinin gene expression occurred earlier under our optimized iPS conditions and the corresponding “neurogenesis burst” associated with proliferative expansion occurred more synchronously, reliably emerging two and three weeks after differentiation. As expected, the expression of mature neuronal markers (i.e., NeuN+/Calbindin+) started at 4-weeks post-differentiation. qPCR microarray, western blot and single cell analyses using high content imaging indicated that 4-week iPS neuronal cultures were non-cycling with decreased expression of cyclin D1 and Ki67. Our data demonstrate that extracellular cues influence the kinetics of neurogenesis models and that feeder layer-free iPSC-derived neurogenesis can be reproducibly miniaturized.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted August 17, 2021.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Extracellular cues accelerate neurogenesis of induced pluripotent stem cell derived neurons
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Extracellular cues accelerate neurogenesis of induced pluripotent stem cell derived neurons
Elizabeth R. Sharlow, Danielle C. Llaneza, Anna J. Mendelson, Garnett A. Mingledorff, Veronica M. Porterfield, Kathryn Salvati, Mark Beenhakker, George S. Bloom, John S. Lazo
bioRxiv 2021.08.17.456634; doi: https://doi.org/10.1101/2021.08.17.456634
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
Extracellular cues accelerate neurogenesis of induced pluripotent stem cell derived neurons
Elizabeth R. Sharlow, Danielle C. Llaneza, Anna J. Mendelson, Garnett A. Mingledorff, Veronica M. Porterfield, Kathryn Salvati, Mark Beenhakker, George S. Bloom, John S. Lazo
bioRxiv 2021.08.17.456634; doi: https://doi.org/10.1101/2021.08.17.456634

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Neuroscience
Subject Areas
All Articles
  • Animal Behavior and Cognition (3688)
  • Biochemistry (7783)
  • Bioengineering (5673)
  • Bioinformatics (21267)
  • Biophysics (10574)
  • Cancer Biology (8170)
  • Cell Biology (11929)
  • Clinical Trials (138)
  • Developmental Biology (6757)
  • Ecology (10394)
  • Epidemiology (2065)
  • Evolutionary Biology (13853)
  • Genetics (9702)
  • Genomics (13063)
  • Immunology (8136)
  • Microbiology (19976)
  • Molecular Biology (7841)
  • Neuroscience (43032)
  • Paleontology (318)
  • Pathology (1278)
  • Pharmacology and Toxicology (2258)
  • Physiology (3350)
  • Plant Biology (7221)
  • Scientific Communication and Education (1311)
  • Synthetic Biology (2000)
  • Systems Biology (5533)
  • Zoology (1127)