Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain.
Författare
Summary, in English
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.
Avdelning/ar
Publiceringsår
2014
Språk
Engelska
Publikation/Tidskrift/Serie
Scientific Reports
Volym
4
Fulltext
Länkar
Dokumenttyp
Artikel i tidskrift
Förlag
Nature Publishing Group
Ämne
- Neurosciences
Status
Published
Projekt
- Cell reprogramming: a new method for generating functional neurons
Forskningsgrupp
- Developmental and Regenerative Neurobiology
ISBN/ISSN/Övrigt
- ISSN: 2045-2322