Neural Stem Cells

What are Neural Stem Cells?

Neural stem cells are a subtype of progenitor cells in the nervous system that can self-renew and give rise to differentiated lineages of neurons and glial cells, including oligodendrocytes and astrocytes.

Neural stem cells are present during neural development and in the adult brain. The stem cell niche architecture allows adult neural stem cells to continuously generate neurons in the brain throughout life. Adult neural stem cells are found mainly in two neurogenic regions: the hippocampus and the subventricular zone (SVZ), as well as in some non-neurogenic regions, such as the spinal cord.

Products

Background

Literature (7)

Cat. No.	Product Name / Activity
2939	A 83-01
	Promotes neural differentiation of hPSCs; selective inhibitor of TGF-βRI, ALK4 and ALK7
7163	Chroman 1
	Highly potent and selective ROCK 2 inhibitor; improves cell survival after cryogenesis
6476	Compound E
	Induces neuronal differentiation; γ-secretase inhibitor
2634	DAPT
	γ-secretase inhibitor; induces neuronal differentiation; blocks Notch signaling
1141	Dibutyryl-cAMP, sodium salt
	Promotes differentiation of hPSCs to dopaminergic neurons; cell-permeable cAMP analog
3093	Dorsomorphin dihydrochloride
	Promotes neural differentiation of hPSCs; also BMP type I receptor inhibitor
0927	Fluoxetine hydrochloride
	5-HT reuptake inhibitor; induces differentiation of neuronal precursors
1099	Forskolin
	Adenylyl cyclase activator; induces neuronal differentiation
4912	GsMTx4
	Neuroprotective compound; enhances neurogenesis in human neural stem cells
7807	Hh-Ag1.5
	Promotes neural differentiation of hPSCs; potent and high affinity Smo receptor agonist
2845	IBMX
	PDE inhibitor; facilitates differentiation of neural progenitor cells
4439	ISX 9
	Neurogenic agent; induces neuronal differentiation of SVZ progenitors and also induces cardiomyogenic differentiation
4888	KHS 101 hydrochloride
	Selective inducer of neuronal differentiation in hippocampal neural progenitors
6053	LDN 193189 dihydrochloride
	Potent and selective ALK2 and ALK3 inhibitor; promotes neural induction of hPSCs
8150	LDN 193189 in solution New
	Sterile-filtered 10 mM solution of LDN 193189 pre-dissolved in water
2864	Metformin hydrochloride
	Activator of LKB1/AMPK; enhances neurogenesis
6668	ML 184
	Promotes NSC proliferation and differentiation; also selective GPR55 agonist
7874	N-Acetylcysteine
	Stimulate neuron differentiation and neuritogenesis of ESCs; also GSH precursor; antioxidant
3854	1-Oleoyl lysophosphatidic acid sodium salt
	LPA₁ and LPA₂ agonist; inhibits differentiation of neural stem cells into neurons
4076	P7C3
	Neuroprotective compound; enhances neurogenesis in vivo
3044	PD 173074
	Selective FGFR1 and -3 inhibitor; inhibits proliferation and differentiation of oligodendrocyte progenitors
3534	PNU 74654
	Promotes neural differentiation of hPSCs; inhibits Wnt signaling
0695	Retinoic acid
	Promotes neural differentiation; endogenous retinoic acid receptor agonist
4366	SAG
	Enhances neuronal differentiation of iPSCs into dopaminergic neurons; Smo agonist
6390	SAG dihydrochloride
	Dihydrochloride salt of SAG; Smo receptor agonist
1614	SB 431542
	Promotes neural differentiation of hPSCs; inhibitor of TGF-βRI, ALK4 and ALK7
3300	SU 5402
	Potent FGFR and VEGFR inhibitor; attenuates integrin β4-induced differentiation of neural stem cells
8093	Tazemetostat New
	Induces neuronal differentiation genes; increases neurite extension. EZH2 inhibitor.
6336	Trazodone hydrochloride
	Enhances neural differentiation; also 5-HT_2A and α₁adrenoceptor antagonist; antidepressant and neuroprotectant
7405	TWS 119
	Induces neuronal differentiation in ESCs; potent GSK3 inhibitor
1254	Y-27632 dihydrochloride
	Selective ROCK inhibitor; part of cocktail used to induce neurons from fibroblasts
7000	Y-27632 in solution New
	Sterile-filtered 10 mM solution of Y-27632 pre-dissolved in water

Neural Stem Cells Differentiation

Neuronal differentiation is guided by key signaling pathways including Wnt, BMP, FGF and retinoic acid (RA) signaling cascades. The generation of neural progenitors is characterized by expression of neuroepithelial markers, such as nestin, PAX6, SOX1, SOX3, PSA-NCAM and MUSASHI-1 in vitro, and the formation of neural rosettes, reminiscent of neural tube initiation in vivo. Various methods have been developed for the conversion of induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) into neuronal lineages, with a view to improving our understanding of neurological diseases and the neurobiological processes involved in development.

Schematic outlining protocol for generation of neurons from hiPSCs

Figure 1. Generation of neurons from hiPSCs. From Qi et al. (2017) Nat.Biotechnol. 35 154. View our stem cells protocols for further information.

Noggin is an endogenous bone morphogenic protein (BMP) antagonist and a known neural-inducing factor. Chambers et al. (2009) reported that dual inhibition of SMAD signaling by Noggin, and the small molecule TGF-β antagonist, SB 431542 (Cat. No. 1614), promotes rapid and complete induction of PAX6+ neuroepithelial cells from hESCs capable of rosette formation. It has subsequently been found that the small molecule ALK2/3 antagonist LDN 193189 (Cat. No. 6053) can substitute Noggin in dual SMAD inhibition for the generation of neural progenitors (NPs). By exposure to small molecule activators of Hedgehog signaling, such as Purmorphamine (Cat. No. 4551), these NPs can be directed to become midbrain floor plate (FP) precursors, characterized by the expression of the marker FOXA2. Activation of Wnt signaling, using the potent GSK-3β inhibitor CHIR 99021 (Cat. No. 4423), promotes the conversion of these FP precursors to a dopamine (DA) neuron fate (Kriks et al., 2011), characterized by the co-expression of FOXA2 and the roof plate marker LMX1A, and in vivo studies showed robust survival and function of the engrafted midbrain DA neurons without neural overgrowth. Li et al. (2011) discovered that combined inhibition of GSK-3β, TGF-β and Notch signaling with CHIR 99021, SB 431542 and the γ-secretase inhibitor Compound E (Cat. No. 6476), respectively, rapidly converts monolayer cultured hESCs to homogenous primitive neuroepithelium within one week. This small molecule cocktail leads to the loss of the pluripotency markers, Oct4 and Nanog with a concurrent increase in PAX6 expression. Expression of Sox2, which is both a pluripotency marker and a marker of neural differentiation, remains unchanged.

Fluorescent images of Cells cultured in Neuronal Media supplemented with TGFb3, cAMP, GDNF (Cat. No. 212-GD, R&D Systems), BDNF (Cat. No. 248-BDB, R&D Systems), N21-MAX (Cat. No. AR008, R&D Systems).

Figure 2. Dopaminergic neurons derived using SB 431542, CHIR 99021, DAPT and Purmorphamine. Cells cultured in Neuronal Media supplemented with TGFβ3 (Cat. No. 8420-B3, R&D Systems), cAMP, GDNF (Cat. No. 212-GD, R&D Systems), BDNF (Cat. No. 248-BDB, R&D Systems), N21-MAX (Cat. No. AR008, R&D Systems). Images courtesy of Kevin Flynn, Bio-Techne.

Cultured DA neurons have the potential to be used as cell therapy for Parkinson's disease (PD). Researchers at Kyoto University (Kikuchi et al., 2017) have generated DA neuron precursors from hiPSCs derived from healthy donors and patients with PD. Their method uses dual SMAD inhibition by LDN 193189 and the TGF-β inhibitor A 83-01 (Cat. No. 2939) for neuronal induction followed by induction of floor plate cells with Purmorphamine, CHIR 99021 and FGF-8. They have shown that dopaminergic progenitors derived in this way survive and function as midbrain DA neurons when transplanted into animal models of PD and that the recipients show increased spontaneous movement following transplantation. The method devised by Kikuchi et al. has been used in a clinical trial for Parkinson's disease at Kyoto University. In the context of stem cell therapy for regenerative medicine, PluriSln 1 (Cat. No. 4847) could be a useful tool as it selectively eliminates undifferentiated hPSCs from culture.

Qi et al. (2017) combined these approaches and devised a protocol using six small molecules for the rapid induction of early-born cortical neurons from hPSCs in 16 days. Their method uses dual SMAD inhibition with LSB, and DAPT (Cat. No. 2634) replaces Compound E as the γ-secretase inhibitor. The tankyrase inhibitor XAV 939 (Cat. No. 3748) is used to activate Wnt signaling in place of CHIR 99021 and additional inhibition of FGF signaling with SU 5402 (Cat. No. 3300) and PD 0325901 (Cat. No. 4192) was also used. Cells cultured in vitro exhibit functional electrophysiological properties by day 16, in the absence of glial cell co-culture. When transplanted into postnatal mouse cortex at day 8 of differentiation the resulting neurons establish long-distance connections (Figure 2).

Literature for Neural Stem Cells

Tocris offers the following scientific literature for Neural Stem Cells to showcase our products. We invite you to request* your copy today!

*Please note that Tocris will only send literature to established scientific business / institute addresses.

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