Abstract
Human pluripotent stem cells (hPSCs ) have the unique potential to form every cell type in the body. This potential provides opportunities for generating human progenitors and other differentiated cell types for understanding human development and for use in cell type-specific therapies . Equally important is the ability to engineer stem cells and their derived progenitors to mimic specific disease models. This chapter will focus on the propagation and characterization of human neural progenitors (hNPs ) derived from hPSCs with a particular focus on engineering hNPs to generate in vitro disease models for human neuro-mitochondrial disorders . We will discuss the methodologies for culturing and characterizing hPSCs and hNPs; and protocols for engineering hNPs by using a novel mitochondrial transfection technology.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Thomson JA, Itskovitz-Eldor J, Shapiro SS et al (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147
Yu J, Vodyanik MA, Smuga-Otto K et al (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318:1917–1920
Takahashi K, Tanabe K, Ohnuki M et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131: 861–872
Shin S, Mitalipova M, Noggle S et al (2006) Long-term proliferation of human embryonic stem cell-derived neuroepithelial cells using defined adherent culture conditions. Stem Cells 24:125–138
Dhara SK, Hasneen K, Machacek DW et al (2008) Human neural progenitor cells derived from embryonic stem cells in feeder-free cultures. Differentiation 76:454–464
Shin S, Dalton S, Stice SL (2005) Human motor neuron differentiation from human embryonic stem cells. Stem Cells Dev 14:266–269
Wilczynska KM, Singh SK, Adams B et al (2009) Nuclear factor I isoforms regulate gene expression during the differentiation of human neural progenitors to astrocytes. Stem Cells 27:1173–1181
Iyer S, Alsayegh K, Abraham S et al (2009) Stem cell-based models and therapies for neurodegenerative diseases. Crit Rev Biomed Eng 37:321–353
Wilson PG, Cherry JJ, Schwamberger S et al (2007) An SMA project report: neural cell-based assays derived from human embryonic stem cells. Stem Cells Dev 16:1027–1041
Khan SM, Bennett JP Jr (2004) Development of mitochondrial gene therapy. J Bioenerg Biomembr 36:387–393
Iyer S, Thomas RR, Portell FR et al (2009) Recombinant mitochondrial transcription factor A with N-terminal mitochondrial transduction domain increases respiration and mitochondrial gene expression. Mitochondrion 9:196–203
Iyer S, Bergquist K, Young K et al (2012) Mitochondrial gene therapy improves respiration, biogenesis, and transcription in G11778A Leber’s hereditary optic neuropathy and T8993G Leigh’s syndrome cells. Hum Gene Ther 23:647–657
Iyer S, Xiao E, Alsayegh K et al (2012) Mitochondrial gene replacement in human pluripotent stem cell-derived neural progenitors. Gene Ther 19:469–475
Acknowledgements
This work was supported in part by start-up funds provided to Dr. Iyer by the Center for the Study of Biological Complexity at Virginia Commonwealth University and a grant from the Presidential Research Innovation Program (PRIP) at Virginia Commonwealth University. The authors would also like to thank Gencia Corporation, Charlottesville for provision of the recombinant human TFAM . Mention of vendor names is for information only and does not imply endorsement.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Rao, R.R., Iyer, S. (2015). Stem Cells, Neural Progenitors, and Engineered Stem Cells. In: Lossi, L., Merighi, A. (eds) Neuronal Cell Death. Methods in Molecular Biology, vol 1254. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2152-2_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2152-2_19
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2151-5
Online ISBN: 978-1-4939-2152-2
eBook Packages: Springer Protocols