Abstract
The human brain and mechanisms underlying its functioning has been a field of intense research due to its complexity, inaccessibility, and the large numbers of debilitating disorders affecting this organ. Model organisms have provided great insight into the functioning of the mammalian brain; however, there exist many features unique to humans which need detailed understanding. In this context, human pluripotent stem cells (HPSCs) have emerged as a promising resource.
In the developing brain, cortical diversification is achieved by neural stem cells/neural progenitor cells (NSCs/NPCs) by altering its potency (from multipotent to unipotent) and differentiation capacity (from neurogenesis to gliogenesis). Recent development in tissue reprogramming allows for derivation of NSCs/NPCs from either healthy control subjects manipulated to carry disease mutations or affected individuals carrying specific disease-causing mutations allowing for detailed evaluation of cellular phenotype, pharmacological manipulation, and/or toxicological screening.
In this chapter, we will discuss HPSC differentiation into neural stem cells (NSCs) and neurons. We will review the mechanism underlying in vivo neural differentiation and methods which recapitulate this in vitro. We describe a method of deriving NSCs and differentiated mature neurons highlighting key steps of the core protocol. We also provide detailed information of the transcription factor and morphogen map of the developing brain which can be used as a guide to derive region- and lineage-specific NSCs and differentiated neurons.
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References
Rakic P (2009) Evolution of the neocortex: a perspective from developmental biology. Nat Rev Neurosci 10(10):724–735
Sasai Y et al (1995) Regulation of neural induction by the Chd and Bmp-4 antagonistic patterning signals in Xenopus. Nature 376(6538):333–336
Fainsod A et al (1997) The dorsalizing and neural inducing gene follistatin is an antagonist of BMP-4. Mech Dev 63(1):39–50
Abranches E et al (2009) Neural differentiation of embryonic stem cells in vitro: a road map to neurogenesis in the embryo. PLoS One 4(7):e6286
Anderson RM et al (2002) Chordin and noggin promote organizing centers of forebrain development in the mouse. Development 129(21):4975–4987
Wurst W, Bally-Cuif L (2001) Neural plate patterning: upstream and downstream of the isthmic organizer. Nat Rev Neurosci 2(2):99–108
Hoch RV, Rubenstein JL, Pleasure S (2009) Genes and signaling events that establish regional patterning of the mammalian forebrain. Semin Cell Dev Biol 20(4):378–386
Efthymiou AG et al (2014) Self-renewal and cell lineage differentiation strategies in human embryonic stem cells and induced pluripotent stem cells. Expert Opin Biol Ther 14(9):1333–1344
Zhang X et al (2010) Pax6 is a human neuroectoderm cell fate determinant. Cell Stem Cell 7(1):90–100
Kunath T et al (2007) FGF stimulation of the Erk1/2 signalling cascade triggers transition of pluripotent embryonic stem cells from self-renewal to lineage commitment. Development 134(16):2895–2902
Eiraku M et al (2008) Self-organized formation of polarized cortical tissues from ESCs and its active manipulation by extrinsic signals. Cell Stem Cell 3(5):519–532
Ho SM, Topol A, Brennand KJ (2015) From “directed differentiation” to “neuronal induction”: modeling neuropsychiatric disease. Biomark Insights 10(Suppl 1):31–41
Li XJ et al (2005) Specification of motoneurons from human embryonic stem cells. Nat Biotechnol 23(2):215–221
Dhara SK, Stice SL (2008) Neural differentiation of human embryonic stem cells. J Cell Biochem 105(3):633–640
Chambers SM et al (2009) Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol 27(3):275–280
Gaspard N, Vanderhaeghen P (2011) Laminar fate specification in the cerebral cortex. F1000 Biol Rep 3:6
Kriks S et al (2011) Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson's disease. Nature 480(7378):547–551
Hoshino M (2006) Molecular mechanisms underlying glutamatergic vs. GABAergic neuronal subtype specification in the cerebellum. Seikagaku 78(2):130–132
Muguruma K, Sasai Y (2012) In vitro recapitulation of neural development using embryonic stem cells: from neurogenesis to histogenesis. Develop Growth Differ 54(3):349–357
Nizzardo M et al (2013) Direct reprogramming of adult somatic cells into other lineages: past evidence and future perspectives. Cell Transplant 22(6):921–944
Liu ML et al (2013) Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons. Nat Commun 4:2183
Pang ZP et al (2011) Induction of human neuronal cells by defined transcription factors. Nature 476(7359):220–223
Heinrich C et al (2010) Directing astroglia from the cerebral cortex into subtype specific functional neurons. PLoS Biol 8(5):e1000373
Victor MB et al (2014) Generation of human striatal neurons by microRNA-dependent direct conversion of fibroblasts. Neuron 84(2):311–323
Caiazzo M et al (2011) Direct generation of functional dopaminergic neurons from mouse and human fibroblasts. Nature 476(7359):224–227
Ring KL et al (2012) Direct reprogramming of mouse and human fibroblasts into multipotent neural stem cells with a single factor. Cell Stem Cell 11(1):100–109
Maroof AM et al (2013) Directed differentiation and functional maturation of cortical interneurons from human embryonic stem cells. Cell Stem Cell 12(5):559–572
Suzuki IK, Vanderhaeghen P (2015) Is this a brain which I see before me? Modeling human neural development with pluripotent stem cells. Development 142(18):3138–3150
Lancaster MA et al (2013) Cerebral organoids model human brain development and microcephaly. Nature 501(7467):373–379
Zirra A et al (2016) Neural Conversion and Patterning of Human Pluripotent Stem Cells: A Developmental Perspective. Stem Cells International 2016:1–14
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Mukherjee, O., Acharya, S., Rao, M. (2019). Making NSC and Neurons from Patient-Derived Tissue Samples. In: Daadi, M. (eds) Neural Stem Cells. Methods in Molecular Biology, vol 1919. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9007-8_2
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DOI: https://doi.org/10.1007/978-1-4939-9007-8_2
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