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Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain
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Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain
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Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain
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Journal Article
Highly efficient generation of induced neurons from human fibroblasts that survive transplantation into the adult rat brain
2014
Overview
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.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 13/100
/ 13/107
/ 38
/ 38/77
/ 42
/ 42/100
/ 64
/ 64/86
/ Animals
/ Basic Helix-Loop-Helix Transcription Factors - genetics
/ Cell Differentiation - physiology
/ Cellular Reprogramming - physiology
/ Female
/ Fetuses
/ Homeodomain Proteins - genetics
/ Humanities and Social Sciences
/ Humans
/ Induced Pluripotent Stem Cells - cytology
/ Medicinska och farmaceutiska grundvetenskaper
/ Microtubule-Associated Proteins - biosynthesis
/ Nerve Tissue Proteins - genetics
/ POU Domain Factors - genetics
/ Rats
/ Rodents
/ Science
/ Smad Proteins - antagonists & inhibitors
/ Transcription Factors - genetics
