NDLI: Efficient derivation of human neuronal progenitors and neurons from pluripotent human embryonic stem cells with small molecule induction.

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Efficient derivation of human neuronal progenitors and neurons from pluripotent human embryonic stem cells with small molecule induction.

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Year: 2011 Issue: 55

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Efficient derivation of human neuronal progenitors and neurons from pluripotent human embryonic stem cells with small molecule induction.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Parsons, Xuejun H. Teng, Yang D. Parsons, James F. Snyder, Evan Y. Smotrich, David B. Moore, Dennis A.
Description	Country affiliation: United States Author Affiliation: Parsons XH ( San Diego Regenerative Medicine Institute, USA. parsons@SDRMI.org)
Abstract	There is a large unfulfilled need for a clinically-suitable human neuronal cell source for repair or regeneration of the damaged central nervous system (CNS) structure and circuitry in today's healthcare industry. Cell-based therapies hold great promise to restore the lost nerve tissue and function for CNS disorders. However, cell therapies based on CNS-derived neural stem cells have encountered supply restriction and difficulty to use in the clinical setting due to their limited expansion ability in culture and failing plasticity after extensive passaging(1-3). Despite some beneficial outcomes, the CNS-derived human neural stem cells (hNSCs) appear to exert their therapeutic effects primarily by their non-neuronal progenies through producing trophic and neuroprotective molecules to rescue the endogenous cells(1-3). Alternatively, pluripotent human embryonic stem cells (hESCs) proffer cures for a wide range of neurological disorders by supplying the diversity of human neuronal cell types in the developing CNS for regeneration(1,4-7). However, how to channel the wide differentiation potential of pluripotent hESCs efficiently and predictably to a desired phenotype has been a major challenge for both developmental study and clinical translation. Conventional approaches rely on multi-lineage inclination of pluripotent cells through spontaneous germ layer differentiation, resulting in inefficient and uncontrollable lineage-commitment that is often followed by phenotypic heterogeneity and instability, hence, a high risk of tumorigenicity(7-10). In addition, undefined foreign/animal biological supplements and/or feeders that have typically been used for the isolation, expansion, and differentiation of hESCs may make direct use of such cell-specialized grafts in patients problematic(11-13). To overcome these obstacles, we have resolved the elements of a defined culture system necessary and sufficient for sustaining the epiblast pluripotence of hESCs, serving as a platform for de novo derivation of clinically-suitable hESCs and effectively directing such hESCs uniformly towards clinically-relevant lineages by small molecules(14) (please see a schematic in Fig. 1). Retinoic acid (RA) does not induce neuronal differentiation of undifferentiated hESCs maintained on feeders(1, 14). And unlike mouse ESCs, treating hESC-differentiated embryoid bodies (EBs) only slightly increases the low yield of neurons(1, 14, 15). However, after screening a variety of small molecules and growth factors, we found that such defined conditions rendered retinoic acid (RA) sufficient to induce the specification of neuroectoderm direct from pluripotent hESCs that further progressed to neuroblasts that generated human neuronal progenitors and neurons in the developing CNS with high efficiency (Fig. 2). We defined conditions for induction of neuroblasts direct from pluripotent hESCs without an intervening multi-lineage embryoid body stage, enabling well-controlled efficient derivation of a large supply of human neuronal cells across the spectrum of developmental stages for cell-based therapeutics.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/3273
Journal	Journal of Visualized Experiments
Issue Number	56
Language	English
Publisher	MyJove Corp.
Publisher Date	2011-10-28
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Cell Culture Techniques Embryonic Stem Cells Cytology Neural Stem Cells Neurons Pluripotent Stem Cells Tretinoin Pharmacology Cell Differentiation Drug Effects Culture Media Nerve Regeneration Research Support, N.i.h., Extramural Video-audio Media
Content Type	Text
Resource Type	Article

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