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Stemdiff neuron differentiation kit

Manufactured by STEMCELL
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The STEMdiff™ Neuron Differentiation Kit is a laboratory product designed to facilitate the differentiation of human pluripotent stem cells into neurons. The kit provides a set of optimized media and reagents to support the stepwise differentiation process.

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Lab products found in correlation

Stemdiff neuron maturation kit, by STEMCELL (2 mentions) Mtesr1 medium, by STEMCELL (2 mentions) Stemdiff neural progenitor medium, by STEMCELL (2 mentions) Brain derived neurotrophic factor (bdnf), by Thermo Fisher Scientific (2 mentions) Neurobasal medium, by Thermo Fisher Scientific (2 mentions) N2 supplement, by Thermo Fisher Scientific (2 mentions) Nestin, by BD (1 mentions) Neural induction medium, by STEMCELL (1 mentions) Matrigel with reduced growth factors, by Thermo Fisher Scientific (1 mentions) H9 wa09 cells, by WiCell (1 mentions) Aggrewell 800 plate, by STEMCELL (1 mentions) Accutase, by STEMCELL (1 mentions) Neural progenitor medium 2, by STEMCELL (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions) Forskolin, by Merck Group (1 mentions) Sb431542, by Bio-Techne (1 mentions) Accutase, by Thermo Fisher Scientific (1 mentions) Dmem f12 medium, by Thermo Fisher Scientific (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Matrigel, by Corning (1 mentions)

4 protocols using stemdiff neuron differentiation kit

1

Tay-Sachs Patient NSCs Neuronalization

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Tay-Sachs patient NSCs were further differentiated into neurons using STEMdiff™ Neuron Differentiation Kit and STEMdiff™ Neuron Maturation Kit (StemCell Technologies) according to the manufacture’s protocol. After 7 days of differentiation of Tay-Sachs NSCs to neuronal precursors in 6- well plate using STEMdiff™ Neuron Differentiation medium, cells were dislodged and seeded into Poly-L-ornithine (PLO)/laminin coated black, clear bottom, tissue-culture treated 96-well plate in STEMdiff™ Neuron Maturation medium for another 7–14 days before downstream application. Immunofluorescence staining of several neuronal markers MAP2 (Cell signaling), beta-III-tubulin (Cell signaling), Neurofilament-L (Cell signaling) as well as Nestin (neural stem cell protein marker, BD Bioscience) were performed to characterize the neuronal cells derived from TSD patient and WT control NSCs.
Vu M., Li R., Baskfield A., Lu B., Farkhondeh A., Gorshkov K., Motabar O., Beers J., Chen G., Zou J., Espejo-Mojica A.J., Rodríguez-López A., Alméciga-Díaz C.J., Barrera L.A., Jiang X., Ory D.S., Marugan J.J, & Zheng W. (2018). Neural stem cells for disease modeling and evaluation of therapeutics for Tay-Sachs disease. Orphanet Journal of Rare Diseases, 13, 152.
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2

Directed Neural Differentiation of H9/WA09 Cells

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H9/WA09 cells (WiCell) were grown on Matrigel with reduced growth factors (ThermoFisher Scientific, #354230) in mTeSR1 medium (STEMCELL Technologies, #85850) at 37°C and 5% CO2. Neural lineage differentiation was performed using STEMdiff™ neuron differentiation kit (STEMCELL Technologies, #08500) and STEMdiff™ neuron maturation kit (STEMCELL Technologies, #08510).ES cells were seeded onto AggreWell800 plates (STEMCELL Technologies, #34811) and fed with neural induction medium (STEMCELL Technologies, #05835) to form uni-sized embryoid bodies (EBs). On day 5, EBs were re-plated onto Matrigel-treated 6-well plates. Neural differentiation started from day 11 to day 16. On day 17, cells were treated with Accutase® (STEMCELL Technologies) and seeded and fed with neuronal maturation medium till days 22–24. RNA samples were collected on days 10, 11, and 22 for deep-sequencing analysis. Cells were also seeded onto multi-chamber slides for immunofluorescence (IF) analysis.
Yang X., Xu B., Mulvey B., Evans M., Jordan S., Wang Y.D., Pagala V., Peng J., Fan Y., Patel A, & Peng J.C. (2019). Differentiation of human pluripotent stem cells into neurons or cortical organoids requires transcriptional co-regulation by UTX and 53BP1. Nature neuroscience, 22(3), 362-373.
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3

Neural Progenitor Cell Differentiation from hiPSCs

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Human neural progenitor cells derived from xcl-1 pluripotent stem cells (male) were obtained from Stem Cell Technologies (cat# 70901) and culture in neural progenitor medium 2 (Stem Cell Technologies, cat# 08560) at 37 °C. Differentiation into neurons was achieved using the STEMdiff™ Neuron Differentiation Kit (Stem Cell Technologies, cat# 08500).
For NPC induction from CRISPRi cells (male), hiPSCs were cultured in DMEM F12 medium (Life Technologies) supplemented with 1X N2 supplement (Life Technologies), 2 mM glutamine (Life Technologies), 100 nM LDN-193189 (Stemgent, 04–0074), 10 μM SB431542 (Tocris, 1254) and 2 μM XAV939 (Stemgent, 04–00046). After 9 days, cells were passaged on Matrigel (Corning) coated dishes using Accutase (Life Technologies) and maintained in STEMdiff™ Neural Progenitor Medium (Stem Cell Technologies). Neuronal differentiation was achieved by culturing NPCs on Matrigel coated dishes in Neurobasal medium supplemented with 1X B27 (Life Technologies), 10 ng/ml BDNF (Peprotech), 10 ng/ml GDNG (Peprotech), ascorbic acid (50 μg/ml, Sigma), 5 M Forskolin (Sigma).
Stefano B.D., Luo E.C., Haggerty C., Aigner S., Charlton J., Brumbaugh J., Ji F., Jiménez I.R., Clowers K.J., Huebner A.J., Clement K., Lipchina I., de Kort M.A., Anselmo A., Pulice J., Gerli M.F., Gu H., Gygi S.P., Sadreyev R.I., Meissner A., Yeo G.W, & Hochedlinger K. (2019). The RNA helicase DDX6 controls cellular plasticity by modulating P-body homeostasis. Cell stem cell, 25(5), 622-638.e13.
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4

Neural Differentiation of Human iPSCs

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Human induced pluripotent stem cell (iPSC) colonies were maintained in mTeSR1 medium (STEMCELL Technologies). Neural progenitor cells (NPCs) were derived from iPSCs using STEMdiff Neuron Differentiation Kit (STEMCELL Technologies) and maintained in STEMdiff Neural Progenitor Medium (STEMCELL Technologies). For the neural differentiation, NPCs were plated at 2 × 104 cells/cm2 onto poly-l-ornithine/laminin coated 15-cm dishes in neural differentiation medium [Neurobasal Medium (Gibco), 1× N-2 supplement (Gibco), 1× B-27 supplement (Gibco), 1× GlutaMAX (Gibco), 0.2 μM l-ascorbic acid (Sigma), 1 μM cAMP (Sigma), 10 ng/ml BDNF (Peprotech), 10 ng/ml GDNF (Peprotech)] supplemented with 0.1 μM Compound E (Millipore) and 5 μM ROCK inhibitor (Millipore). Neural cultures were maintained in neural differentiation medium for 1 month before collection. For the iPSC sample collection, cyclohexmide was added to the medium to a final concentration of 100 μg/ml and cells were incubated at 37°C for 1 min. The cells were subsequently washed with ice-cold PBS containing 100 μg/ml CHX, collected by scraping from the dish, pelleted by centrifugation at 15 000g 4°C for 3 min, snap-frozen in liquid nitrogen, and immediately stored at −80°C.
Liu B., Molinaro G., Shu H., Stackpole E.E., Huber K.M, & Richter J.D. (2018). Optimization of ribosome profiling using low-input brain tissue from fragile X syndrome model mice. Nucleic Acids Research, 47(5), e25.
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