Capture of Neuroepithelial-Like Stem Cells from Pluripotent Stem Cells Provides a Versatile System for In Vitro Production of Human Neurons

Abstract

Human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) provide new prospects for studying human neurodevelopment and modeling neurological disease. In particular, iPSC-derived neural cells permit a direct comparison of disease-relevant molecular pathways in neurons and glia derived from patients and healthy individuals. A prerequisite for such comparative studies are robust protocols that efficiently yield standardized populations of neural cell types. Here we show that long-term self-renewing neuroepithelial-like stem cells (lt-NES cells) derived from 3 hESC and 6 iPSC lines in two independent laboratories exhibit consistent characteristics including i) continuous expandability in the presence of FGF2 and EGF; ii) stable neuronal and glial differentiation competence; iii) characteristic transcription factor profile; iv) hindbrain specification amenable to regional patterning; v) capacity to generate functionally mature human neurons. We further show that lt-NES cells are developmentally distinct from fetal tissue-derived radial glia-like stem cells. We propose that lt-NES cells provide an interesting tool for studying human neurodevelopment and may serve as a standard system to facilitate comparative analyses of hESC and hiPSC-derived neural cells from control and diseased genetic backgrounds.

Treatment of a Mouse Model of Spinal Cord Injury by Transplantation of Human Induced Pluripotent Stem Cell-Derived Long-Term Self-Renewing Neuroepithelial-Like Stem Cells

“…Uninfected and infected hiPS-lt-NES cells were induced to differentiate by removal of growth factors and cultured in the presence of 1% FBS for 4 weeks. Both hiPS-lt-NES cell population differentiated into large and small numbers of Tuj1-positive neurons and GFAP-positive astrocytes, respectively, as observed in our previous study [11] (Fig. 1D).…”

“…We have previously shown that hiPS-NES cells, which are reliably derived from different hiPS cell lines, exhibit consistent characteristics such as continuous expandability, stable neuronal and glial differentiation, and the capacity to generate functional mature human neurons [11]. hiPS-lt-NES cells can be expanded in the presence of FGF2 and EGF in monolayer culture, and express the NSC markers Sox2 and Nestin (Fig.…”

“…hsp-NSCs and hiPS-lt-NES cells were established and maintained as described previously [10,11,16]. The hsp-NS cell line CB660sp and hiPS-lt-NES cell line AF22 were used in the present study.…”

“…We used a recently developed protocol for the generation of long-term self-renewing neuroepithelial-like stem (lt-NES) cells, which satisfy the criteria to be defined as NSCs, from several different lines of hESCs and hiPSCs [10,11]. hiPSCderived lt-NES (hiPS-lt-NES) cells exhibit consistent characteristics such as continuous expandability, stable neuronal and glial differentiation ability, and the capacity to generate functional mature neurons in monolayer culture.…”

Genome Editing in Neuroepithelial Stem Cells to Generate Human Neurons with High Adenosine-Releasing Capacity

“…In this context, we have recently described a population of hESC‐derived long‐term self‐renewing neuroepithelial stem cells (lt‐NES) 22, 23. Similar to pluripotent stem cells, this population exhibits strong self‐renewal capacity enabling genetic modification, subsequent clonal selection and expansion at a scale sufficient for potential therapy.…”