Derivation of High Purity Neuronal Progenitors from Human Embryonic Stem Cells

Abstract

The availability of human neuronal progenitors (hNPs) in high purity would greatly facilitate neuronal drug discovery and developmental studies, as well as cell replacement strategies for neurodegenerative diseases and conditions, such as spinal cord injury, stroke, Parkinson's disease, Alzheimer's disease, and Huntington's disease. Here we describe for the first time a method for producing hNPs in large quantity and high purity from human embryonic stem cells (hESCs) in feeder-free conditions, without the use of exogenous noggin, sonic hedgehog or analogs, rendering the process clinically compliant. The resulting population displays characteristic neuronal-specific markers. When allowed to spontaneously differentiate into neuronal subtypes in vitro, cholinergic, serotonergic, dopaminergic and/or noradrenergic, and medium spiny striatal neurons were observed. When transplanted into the injured spinal cord the hNPs survived, integrated into host tissue, and matured into a variety of neuronal subtypes. Our method of deriving neuronal progenitors from hESCs renders the process amenable to therapeutic and commercial use.

RETRACTED ARTICLE: Analyzing time-series microarray data reveals key genes in spinal cord injury

“…Both two pathways have been reported to be involved in SCI [28][29][30][31][32]. Wnt proteins are a large family of factors that play important roles in embryonic development, including axis patterning, proliferation, cell fate determination, and axon development.…”

Direct Conversion of Human Fibroblasts into Neuronal Restricted Progenitors

“…In previous efforts, NRP-like cells have been induced from human ES cells (28). The small amount of residual pluripotent stem cells among the induced cells can be a safety concern because of the risk of their tumorigenic tendency (29,30).…”

Neural differentiation from pluripotent stem cells: The role of natural and synthetic extracellular matrix

“…Robust neural differentiation has been observed from various hESC and hiPSC lines, although variations among cell lines exist (Table 3) [12,43,44] . The differentiation of hPSCs into NPCs has been performed either by monolayer induction or by the formation of EBs in suspension, with inducing factors including RA, FGF-2, EGF and Shh, etc [45][46][47][48][49] . Recently, the synergistic induction using two inhibitors of SMAD signaling, noggin and SB431642, resulted in efficient neural differentiation for various hPSC lines [12,50] .…”

“…However, the populations obtained in different studies had different potential to differentiate into mature neuronal types. For example, FGF-2/EGF expanded hiPSCderived NSCs showed a high tendency to differentiate into γ-aminobutyric acid neurons while RA/FGF-2 induced hESC-derived NPCs differentiated easily into motor neurons [46,47] . Specific neuronal cell types are required for treating particular neurological diseases.…”

“…However, the cell engraftment and in vivo maturation are yet to be improved. Transplantations of NPCs derived from hiPSCs for treating other neurological diseases such as ALS and muscular atrophy have also been demonstrated in proof-of-principle studies [47,57] . The neural progenitors survived and engrafted in vivo, and the nestin-positive cells differentiated into neuronal phenotype and motoneuron-like structure in both wild-type rats and the ALS rats harboring a mutated human SOD1 (G93A) gene [57] .…”