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Stem Cells Transl Med. 2017 Apr;6(4):1217-1226. doi: 10.1002/sctm.16-0371. Epub 2017 Feb 18.

Highly Efficient Neural Conversion of Human Pluripotent Stem Cells in Adherent and Animal-Free Conditions.

Author information

1
Stem Cells Therapies in Neurodegenerative Diseases Lab.
2
National Stem Cell Bank-Valencia Node, Biomolecular and Bioinformatics Resources Platform PRB2,ISCIII.
3
CABIMER (Centro Andaluz de Biología Molecular y Medicina Regenerativa), Avda. Americo Vespucio s/n, Parque Científico y Tecnológico Cartuja, Sevilla, Spain.
4
Spebo Medical, Leskovac, Serbia.
5
Instituto de Biomedicina de Sevilla (IBiS) and Departamento de Fisiología Médica y Biofísica, Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, Seville, Spain.
6
Department of Neuroscience, Institute of Experimental Medicine, Academy of Science of the Czech Republic, Prague, Czech Republic.
7
Neuronal And Tissue Regeneration Lab, Research Center "Principe Felipe", Valencia, Spain.
8
Faculty of Medical Sciences, Human Genetics Department, University of Kragujevac, Serbia.

Abstract

Neural differentiation of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) can produce a valuable and robust source of human neural cell subtypes, holding great promise for the study of neurogenesis and development, and for treating neurological diseases. However, current hESCs and hiPSCs neural differentiation protocols require either animal factors or embryoid body formation, which decreases efficiency and yield, and strongly limits medical applications. Here we develop a simple, animal-free protocol for neural conversion of both hESCs and hiPSCs in adherent culture conditions. A simple medium formula including insulin induces the direct conversion of >98% of hESCs and hiPSCs into expandable, transplantable, and functional neural progenitors with neural rosette characteristics. Further differentiation of neural progenitors into dopaminergic and spinal motoneurons as well as astrocytes and oligodendrocytes indicates that these neural progenitors retain responsiveness to instructive cues revealing the robust applicability of the protocol in the treatment of different neurodegenerative diseases. The fact that this protocol includes animal-free medium and human extracellular matrix components avoiding embryoid bodies makes this protocol suitable for the use in clinic. Stem Cells Translational Medicine 2017;6:1217-1226.

KEYWORDS:

Cellular therapy; Clinical translation; Differentiation; Embryonic stem cells; Induced pluripotent stem cells; Neural differentiation; Pluripotent stem cells

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