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Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain.

Pereira, Maria J M LU ; Pfisterer, Ulrich LU ; Rylander, Daniella LU ; Torper, Olof LU ; Lau, Shong LU ; Lundblad, Martin LU ; Grealish, Shane LU and Parmar, Malin LU (2014) In Scientific Reports 4.
Abstract
Induced neurons (iNs) offer a novel source of human neurons that can be explored for applications of disease modelling, diagnostics, drug screening and cell replacement therapy. Here we present a protocol for highly efficient generation of functional iNs from fetal human fibroblasts, and also demonstrate the ability of these converted human iNs (hiNs) to survive transplantation and maintain their phenotype in the adult rat brain. The protocol encompasses a delay in transgene activation after viral transduction that resulted in a significant increase in conversion efficiency. Combining this approach with treatment of small molecules that inhibit SMAD signalling and activate WNT signalling provides a further increase in the conversion... (More)
Induced neurons (iNs) offer a novel source of human neurons that can be explored for applications of disease modelling, diagnostics, drug screening and cell replacement therapy. Here we present a protocol for highly efficient generation of functional iNs from fetal human fibroblasts, and also demonstrate the ability of these converted human iNs (hiNs) to survive transplantation and maintain their phenotype in the adult rat brain. The protocol encompasses a delay in transgene activation after viral transduction that resulted in a significant increase in conversion efficiency. Combining this approach with treatment of small molecules that inhibit SMAD signalling and activate WNT signalling provides a further increase in the conversion efficiency and neuronal purity, resulting in a protocol that provides a highly efficient method for the generation of large numbers of functional and transplantable iNs from human fibroblasts without the use of a selection step. When transplanting the converted neurons from different stages of in vitro culture into the brain of adult rats, we observed robust survival and maintenance of neuronal identity four weeks post-transplantation. Interestingly, the positive effect of small molecule treatment observed in vitro did not result in a higher yield of iNs surviving transplantation. (Less)
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organization
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publication status
published
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in
Scientific Reports
volume
4
publisher
Nature Publishing Group
external identifiers
  • pmid:25208484
  • wos:000341934500004
  • scopus:84915734112
ISSN
2045-2322
DOI
10.1038/srep06330
language
English
LU publication?
yes
id
6cd741e6-ada8-4ffd-92d2-75fe2d28c32b (old id 4691910)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/25208484?dopt=Abstract
date added to LUP
2014-10-04 21:52:06
date last changed
2017-10-22 04:21:19
@article{6cd741e6-ada8-4ffd-92d2-75fe2d28c32b,
  abstract     = {Induced neurons (iNs) offer a novel source of human neurons that can be explored for applications of disease modelling, diagnostics, drug screening and cell replacement therapy. Here we present a protocol for highly efficient generation of functional iNs from fetal human fibroblasts, and also demonstrate the ability of these converted human iNs (hiNs) to survive transplantation and maintain their phenotype in the adult rat brain. The protocol encompasses a delay in transgene activation after viral transduction that resulted in a significant increase in conversion efficiency. Combining this approach with treatment of small molecules that inhibit SMAD signalling and activate WNT signalling provides a further increase in the conversion efficiency and neuronal purity, resulting in a protocol that provides a highly efficient method for the generation of large numbers of functional and transplantable iNs from human fibroblasts without the use of a selection step. When transplanting the converted neurons from different stages of in vitro culture into the brain of adult rats, we observed robust survival and maintenance of neuronal identity four weeks post-transplantation. Interestingly, the positive effect of small molecule treatment observed in vitro did not result in a higher yield of iNs surviving transplantation.},
  articleno    = {6330},
  author       = {Pereira, Maria J M and Pfisterer, Ulrich and Rylander, Daniella and Torper, Olof and Lau, Shong and Lundblad, Martin and Grealish, Shane and Parmar, Malin},
  issn         = {2045-2322},
  language     = {eng},
  publisher    = {Nature Publishing Group},
  series       = {Scientific Reports},
  title        = {Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain.},
  url          = {http://dx.doi.org/10.1038/srep06330},
  volume       = {4},
  year         = {2014},
}