Dopaminergic Differentiation of Neural Stem Cells

Neural stem cells (NSCs) were maintained in a growth medium comprised of Neurobasal medium (Gibco), B-27 (Gibco), non-essential amino acid (Sigma-Aldrich), GlutaMAX (Gibco), and 10 ng/mL of fibroblast growth factor (FGF). Differentiation into a dopaminergic phenotype was carried out over 10 days in medium comprised of Neurobasal medium, 1X B27 supplements, 1X non-essential amino acids, 20 ng/mL brain-derived neurotrophic factor (BDNF), 20 ng/mL glial-derived neurotrophic factor (GDNF), and 20 ng/mL TGFb3.^{101 (link)} All growth factors were purchased from Peprotech (Rocky Hill, NJ). NSCs were grown in plates/flasks coated with Geltrex ™ (ThermoFisher Scientific, Carlsbad, CA). 50% of the differentiation media was replaced with fresh media every 72 hours.

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Moritz A.E., Free R.B., Weiner W.S., Akano E.O., Gandhi D., Abramyan A., Keck T.M., Ferrer M., Hu X., Southall N., Steiner J., Aubé J., Shi L., Frankowski K.J, & Sibley D.R. (2020). Discovery, Optimization and Characterization of ML417: A Novel and Highly Selective D3 Dopamine Receptor Agonist. Journal of medicinal chemistry, 63(10), 5526-5567.

Publication 2020

Neurobasal medium non-essential amino acid GlutaMAX non-essential amino acids Geltrex ™

Brain derived neurotrophic factor Dopaminergic Essential amino acids Fibroblast growth factor 10 Glial derived neurotrophic factor Growth factors Neural stem cells Phenotype Supplements Tgfb3

Corresponding Organization : University of North Carolina at Chapel Hill

Other organizations : National Institute on Drug Abuse, Rowan University, National Center for Advancing Translational Sciences

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Variable analysis

independent variables

Differentiation medium composition
Growth medium composition

dependent variables

Differentiation into dopaminergic phenotype

control variables

Neurobasal medium
B-27 supplements
Non-essential amino acids
GlutaMAX
Geltrex coating on plates/flasks

positive controls

10 ng/mL fibroblast growth factor (FGF) in growth medium

negative controls

Not explicitly mentioned

Annotations

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