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Chir 99021

Chir 99021 is a highly selective and potent inhibitor of glycogen synthase kinase-3 (GSK-3), a key regulator of diverse cellular processes.
It has been widely used in research to investigate the role of GSK-3 in various biological systems, including stem cell biology, neurodegeneration, and cancer.
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Most cited protocols related to «Chir 99021»

1
Differentiation and Characterization of Midbrain Dopaminergic Neurons
Cited 143 times
Human ESC (H9, H1) and iPSC lines (2C6 and SeV6) were subjected to a modified dual SMAD-inhibition13 (link) based FP induction12 (link) protocol. Exposure to SHH C25II, Purmorphamine, FGF8 and CHIR99021 were optimized for midbrain FP and DA neuron yield (see Figure 1d). Following FP induction, further maturation was carried out in Neurobasal/B27 medium supplemented with AA, BDNF, GDNF, TGFβ3 and dbcAMP (see full methods for details). The resulting DA neurons were subjected to extensive phenotypic characterization via immunocytochemistry, qRT-PCR, gene expression profiling, HPLC analysis for DA and in vitro electrophysiological recordings. In vivo studies were performed in 6-hydroxydopamine lesioned, hemiparkinsonian rodents (adult NOD-SCID IL2Rgc mice and Sprague Dawley rats) as well as in two adult rhesus monkeys treated with carotid injections of MPTP. DA neurons were injected stereotactically in the striata of the animals (150 × 103 cells in mice, 250 × 103 cells in rats) and a total of 7.5 × 106 cells (distributed in 6 tracts; 3 on each side of brain) in monkeys. Behavioral assays were performed at monthly intervals post grafting, including amphetamine mediated rotational analysis as well as a test for focal akinesia (“stepping test”) and forelimb use (cylinder test). Rats and mice were sacrificed at 18–20 weeks and the primates at 1 month post grafting. Characterization of the grafts was performed via stereological analyses of cell numbers and graft volumes and comprehensive immunohistochemistry.
Kriks S., Shim J.W., Piao J., Ganat Y.M., Wakeman D.R., Xie Z., Carrillo-Reid L., Auyeung G., Antonacci C., Buch A., Yang L., Beal M.F., Surmeier D.J., Kordower J.H., Tabar V, & Studer L. (2011). Floor plate-derived dopamine neurons from hESCs efficiently engraft in animal models of PD. Nature, 480(7378), 547-551.
Publication 2011
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine Adult Amphetamine Animals Biological Assay Brain Bucladesine Carotid Arteries Cells Chir 99021 FGF8 protein, human Forelimb Glial Cell Line-Derived Neurotrophic Factor Grafts High-Performance Liquid Chromatographies Homo sapiens Hydroxydopamine Immunocytochemistry Immunohistochemistry Induced Pluripotent Stem Cells Macaca mulatta Mesencephalon Mice, Inbred NOD Monkeys Mus Neurons Phenotype Primates purmorphamine Rats, Sprague-Dawley Rattus Rodent SCID Mice Step Test Striatum, Corpus
2
Directed Differentiation of hPSCs into SC-β Cells
Cited 63 times

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Publication 2014
Betacellulin Cell Lines Cells Chir 99021 Differentiations, Cell Human Embryonic Stem Cells Human Induced Pluripotent Stem Cells Humidity LDN 193189 Population Group Stem Cells Y 27632
3
Neural Induction and Nociceptor Differentiation
Cited 58 times
Neural induction was performed as previously reported6 (link). Briefly, cells were rendered to single cells using accutase plated on gelatin for 30 minutes to remove MEFs. Non-adherent cells were collected and plated on matrigel treated dishes at a density of 20-40,000 cells/cm2 (link) in the presence of MEF-conditioned hESC media containing 10 ng/ml FGF-2 and 10 μM Y-27632. Neural differentiation was initiated when the cells were confluent using KSR media containing 820 ml of Knockout DMEM, 150 ml Knockout Serum Replacement, 1 mM L-glutamine, 100 μM MEM non-essential amino acids, and 0.1 mM β-mercaptoethanol. To inhibit SMAD signaling, 100nM LDN-193189 and 10 μM SB431542 were added on days 0 through 5. Cells were fed daily, and N2 media was added in increasing 25% increments every other day starting on day 4 (100% N2 on day 10). Nociceptor induction was initiated with the addition of the three inhibitors 3 μM CHIR99021, 10 μM SU5402, 10 μM DAPT on days 2 through 10. Cell passage to lower density can promote maturation of SOX10+ progenitors, and long-term culture media consisted of N2 containing 25ng/ml human-b-NGF, BDNF, and GDNF.
Chambers S.M., Qi Y., Mica Y., Lee G., Zhang X.J., Niu L., Bilsland J., Cao L., Stevens E., Whiting P., Shi S.H, & Studer L. (2012). Combined small molecule inhibition accelerates developmental timing and converts human pluripotent stem cells into nociceptors. Nature biotechnology, 30(7), 715-720.
Publication 2012
1,2-dilinolenoyl-3-(4-aminobutyryl)propane-1,2,3-triol 2-Mercaptoethanol 4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide accutase Amino Acids, Essential Cardiac Arrest Cells Chir 99021 Culture Media Culture Media, Conditioned Fibroblast Growth Factor 2 Gelatins Glial Cell Line-Derived Neurotrophic Factor Glutamine Homo sapiens Human Embryonic Stem Cells Hyperostosis, Diffuse Idiopathic Skeletal inhibitors LDN 193189 matrigel Nervousness Nociceptors Serum SOX10 Transcription Factor SU 5402 Y 27632
4
Isolation and Culture of Airway Stem Cells
Cited 29 times

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Publication 2016
Bronchi Cell Culture Techniques Cells Chir 99021 Culture Media Culture Media, Conditioned Culture Techniques Homo sapiens isolation Laminin Mus Stem Cells Tissues Trachea Trypsin
5
Directed Differentiation of hPSCs
Cited 27 times

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Publication 2014
A-83-01 activin A Bone Morphogenetic Protein 4 Chir 99021 Collagenase Fibroblast Growth Factor 2 FN1 protein, human matrigel PI103

Most recents protocols related to «Chir 99021»

1
Establishment and Characterization of PDTO
Tumor pieces will be dissociated enzymatically and mechanically to obtain isolated cells or small cell clusters (Fig. 2). Cells will be embedded in an extracellular matrix (growth factor-reduced Matrigel or BME II) and cultured in a medium supplemented with growth factors and signal pathway inhibitors [Advanced DMEM (Gibco) supplemented with 100 UI/mL of penicillin and streptomycin (Gibco), 1% GlutaMAX (Gibco), 1X B27 (Gibco), 1.25 mM NAC (Sigma-Aldrich), 50 ng/mL EGF (PeproTech), 10 ng/mL FGF-10 (PeproTech), 5 ng/mL FGF-b (PeproTech), 500 nM A-83-01 (PeproTech), 10 μM Y27632 (Interchim), 10 mM Nicotinamide (Sigma-Aldrich), 1 μM PGE2 (PeproTech), 1 μM Forskolin (Peprotech), 0.3 μM CHIR99021 (Biogems), 100 μg/mL Primocin (InvivoGen), 50% Wnt3a, RSPO3, Noggin-conditioned media (L-WRN, ATCC), and 10% RSPO1-conditioned media (Cultrex HA-R-Spondin-1-Fc 293 T, Amsbio)]. Culture medium will be changed twice a week. Once formed, PDTO will be dissociated and reseeded to amplify them for experimental purposes. Cryovials will be prepared at regular intervals by dissociating and resuspending PDTO in Recovery Cell Culture Freezing Medium (Gibco) prior to be biobanked in liquid nitrogen. It should be noted that PDTO line will be considered as established when it will be maintained for more than 3 passages. For each PDTO line, samples will be kept frozen for DNA/RNA/protein analysis and others will be embedded in paraffin for histopathological analysis. This will allow comparisons between the characteristics of the PDTO and the tumor from which they are derived in order to validate their correspondence.

Establishment and characterization of PDTO derived from HNSCC and evaluation of response to treatments to assess its predictive value

Perréard M., Florent R., Divoux J., Grellard J.M., Lequesne J., Briand M., Clarisse B., Rousseau N., Lebreton E., Dubois B., Harter V., Lasne-Cardon A., Drouet J., Johnson A., Le Page A.L., Bazille C., Jeanne C., Figeac M., Goardon N., Vaur D., Micault E., Humbert M., Thariat J., Babin E., Poulain L., Weiswald L.B, & Bastit V. (2023). ORGAVADS: establishment of tumor organoids from head and neck squamous cell carcinoma to assess their response to innovative therapies. BMC Cancer, 23, 223.
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Publication 2023
Cell Culture Techniques Cells Chir 99021 Colforsin Culture Media Culture Media, Conditioned Dinoprostone Extracellular Matrix Freezing Growth Factor HSP40 Heat-Shock Proteins inhibitors matrigel Neoplasms Niacinamide Nitrogen noggin protein Paraffin Embedding Penicillins Signal Pathways Squamous Cell Carcinoma of the Head and Neck Streptomycin Y 27632
2
Cryopreserved Hepatocyte Expansion Protocol
The cryopreserved human hepatocytes were plated on collagen type I-coated (Rat-tail collagen type I, Gibco) plate at a density of 1 × 105/cm2 in a modified liver expansion medium containing William’s E Medium supplemented with 2% B27 (without VA), 1% PS (All from Invitrogen), 50 ng/mL EGF, 20 ng/mL HGF (Both from Peprotech), 200 μM 2-phospho-L-ascorbic acid (pVc), 3 μM CHIR99021, 5 μM SB431542, 5 μM Lysophosphatidic acid (LPA), 0.5 μM Sphingosine-1-phosphate (S1P) (All from MCE), and 1% fetal bovine serum (FBS) (Gibco, US). After seeding for 7–10 days, expanded cells were passaged at a ratio of 1:2 after dissociation with Accutase (eBioscience). The expansion medium was changed every day. Hepatocyte donor information is shown in Table 1.
Shi J., Li G., Yuan X., Wang Y., Gong M., Li C., Ge X, & Lu S. (2023). Exploration and verification of COVID-19-related hub genes in liver physiological and pathological regeneration. Frontiers in Bioengineering and Biotechnology, 11, 1135997.
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Publication 2023
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide accutase Ascorbic Acid Cells Chir 99021 Collagen Type I Fetal Bovine Serum Hepatocyte Homo sapiens Liver lysophosphatidic acid sphingosine 1-phosphate Tail Tissue Donors
3
Differentiation of hiPSCs into Endothelial Cells
We differentiated hiPSCs into ECs according to the protocol previously described by Patsch et al. (36 (link)) and then adapted for differentiation in 3D culture. In brief, hiPSCs were dissociated from 10-cm plates using ReLeSR (StemCell Technologies) according to the manufacturer’s instructions. Cells were seeded into 125-ml Erlenmeyer flasks at 200,000 cells/ml in 20 ml of StemScale media (Thermo Fisher Scientific) containing 10 μM Y-27632 and then placed on a plate shaker rotating at 70 rpm. Media were changed to fresh StemScale media the next day to remove the ROCK inhibitor. EC differentiation began 4 days after seeding, when spheroids reached approximately 300 μm in diameter. On day 0 of differentiation, media were changed to N2B27 medium containing 7 μM CHIR99021 and BMP-4 (25 μg/ml) and incubated for 3 days without media change. On day 3 of differentiation, media were changed to StemPro-34 containing GlutaMAX (Thermo Fisher Scientific) with 2 μM forskolin (Selleck Chemicals) and VEGF (200 ng/ml) (PeproTech). This same media composition with StemPro-34, forskolin, and VEGF was changed fresh daily for 3 days through day 5 of differentiation. On day 6 of differentiation, spheroids were dissociated using 0.5% trypsin-EDTA diluted 1:3 in phosphate-buffered saline (PBS) for ~5 min. CD144+ ECs were sorted using magnetic microbeads using magnetic-activated cell sorting columns (Miltenyi Biotec) according to the manufacturer’s instructions. Sorted CD144+ ECs were replated on Geltrex-coated 10-cm plates and allowed to proliferate for 3 to 7 days in StemPro-34 containing GlutaMAX and VEGF (50 ng/ml) and then redissociated with trypsin-EDTA and replated again onto Geltrex-coated plates for assay.
Lin Z., Garbern J.C., Liu R., Li Q., Mancheño Juncosa E., Elwell H.L., Sokol M., Aoyama J., Deumer U.S., Hsiao E., Sheng H., Lee R.T, & Liu J. (2023). Tissue-embedded stretchable nanoelectronics reveal endothelial cell–mediated electrical maturation of human 3D cardiac microtissues. Science Advances, 9(10), eade8513.
Publication 2023
Biological Assay BMP4 protein, human Cells Chir 99021 Colforsin Edetic Acid Human Induced Pluripotent Stem Cells Microspheres Phosphates Saline Solution Stem Cells Trypsin Vascular Endothelial Growth Factors Y 27632
4
Cardiomyocyte Differentiation from hiPSCs
hiPSCs were seeded into 10-cm tissue culture plates and then allowed to grow for ~4 days until the cells became ~70% confluent. CMs were differentiated from iPSCs as previously described (79 (link)) [protocol originally adapted from Lian et al. (35 (link))]. Media were changed to RPMI 1640 + B27 + ascorbic acid (RPMI/B27/AA) containing the GSK3 inhibitor CHIR99021 (Selleck Chemicals) (6 μM) on day 0 of CM differentiation. On day 2 of differentiation, media were changed to RPMI/B27/AA containing the Wnt antagonist IWP-4 (Reprocell) (5 μM). On day 4 of differentiation, media were changed to fresh RPMI/B27/AA, and on day 7 of differentiation, basal media were changed to RPMI/B27 + insulin (10 μg/ml), with media subsequently changed every 2 to 3 days with RPMI/B27/insulin. Beating of CMs was usually observed between days 7 and 9 of differentiation. CMs were dissociated using the STEMdiff Cardiomyocyte Dissociation Kit (StemCell Technologies) and replated onto Geltrex-coated plates for further assay.
Lin Z., Garbern J.C., Liu R., Li Q., Mancheño Juncosa E., Elwell H.L., Sokol M., Aoyama J., Deumer U.S., Hsiao E., Sheng H., Lee R.T, & Liu J. (2023). Tissue-embedded stretchable nanoelectronics reveal endothelial cell–mediated electrical maturation of human 3D cardiac microtissues. Science Advances, 9(10), eade8513.
Publication 2023
Ascorbic Acid Biological Assay Cells Chir 99021 Glycogen Synthase Kinase 3 Human Induced Pluripotent Stem Cells Induced Pluripotent Stem Cells Insulin Melia azedarach Myocytes, Cardiac Stem Cells Tissues
5
Cardiomyocyte Differentiation from hiPSCs
The hiPSC line was adopted in this study after obtaining them from the Stanford Cardiovascular Institute (SCVI) Biobank, Stanford University. It was generated thorough reprograming of peripheral blood mononuclear cells (PBMCs) from an anonymous healthy individual with Sendai virus. The pluripotent cell lines were grown in Matrigel (Corning, CA)-coated 12-well plates in Essential 8™ Medium (Thermo Fisher Scientific, MA) at 37°C in 5% CO2 in compressed air and high humidity. Cardiomyocyte differentiation was conducted using a monolayer differentiation chemically defined method (Burridge et al., 2014 (link)). Briefly, iPSCs were kept in culture until 80%–90% confluence. For the differentiation, iPSCs were treated with 6 µM CHIR99021 in RPMI + B27 (minus insulin) for 2 days, fresh RPMI + B27 (minus insulin) for 1 day, followed by 5 µM IWR-1 treatment for 2 days, and then fresh RPMI + B27 (minus insulin) for another 2 days. Afterward, the cells will be supplied with fresh RPMI + B27 every other day. Beating cardiomyocytes will normally appear after 9–10 days, and the cells can be further treated with glucose free RPMI + B27 for 2–3 rounds.
Zhang P., Liu Y., Li C., Stine L.D., Wang P.H., Turnbull M.W., Wu H, & Liu Q. (2023). Ectopic expression of SARS-CoV-2 S and ORF-9B proteins alters metabolic profiles and impairs contractile function in cardiomyocytes. Frontiers in Cell and Developmental Biology, 11, 1110271.
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Publication 2023
Cardiovascular System Cell Lines Cells Chir 99021 Glucose Human Induced Pluripotent Stem Cells Humidity Induced Pluripotent Stem Cells Insulin matrigel Myocytes, Cardiac PBMC Peripheral Blood Mononuclear Cells Sendai virus

Top products related to «Chir 99021»

1
Chir99021 by Selleck Chemicals
Sourced in United States, China, United Kingdom, Canada
CHIR99021 is a small molecule chemical compound used in laboratory research. It functions as a selective inhibitor of the glycogen synthase kinase-3 (GSK-3) enzyme.
2
Chir99021 by Bio-Techne
Sourced in United Kingdom, United States, Germany
CHIR99021 is a small molecule compound that functions as a glycogen synthase kinase-3 (GSK-3) inhibitor. It is commonly used in cell culture and stem cell research applications.
3
Glutamax by Thermo Fisher Scientific
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GlutaMAX is a chemically defined, L-glutamine substitute for cell culture media. It is a stable source of L-glutamine that does not degrade over time like L-glutamine. GlutaMAX helps maintain consistent cell growth and performance in cell culture applications.
4
Chir99021 by Merck Group
Sourced in United States, Germany, Canada, United Kingdom, France
CHIR99021 is a small molecule that functions as a highly selective and potent inhibitor of glycogen synthase kinase-3 (GSK-3). It is commonly used as a laboratory reagent in cell culture and biochemical studies.
5
Non essential amino acids (neaa) by Thermo Fisher Scientific
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Non-essential amino acids are a group of amino acids that can be synthesized by the human body and are not required to be obtained through diet. These amino acids play a fundamental role in various biological processes, including protein synthesis and cellular function.
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Penicillin streptomycin by Thermo Fisher Scientific
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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Dmem f12 by Thermo Fisher Scientific
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DMEM/F12 is a cell culture medium developed by Thermo Fisher Scientific. It is a balanced salt solution that provides nutrients and growth factors essential for the cultivation of a variety of cell types, including adherent and suspension cells. The medium is formulated to support the proliferation and maintenance of cells in vitro.
8
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
9
B27 supplement by Thermo Fisher Scientific
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B27 supplement is a serum-free and animal component-free cell culture supplement developed by Thermo Fisher Scientific. It is designed to promote the growth and survival of diverse cell types, including neurons, embryonic stem cells, and other sensitive cell lines. The core function of B27 supplement is to provide a defined, optimized combination of vitamins, antioxidants, and other essential components to support cell culture applications.
10
L glutamine by Thermo Fisher Scientific
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L-glutamine is an amino acid that is commonly used as a dietary supplement and in cell culture media. It serves as a source of nitrogen and supports cellular growth and metabolism.

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