CHIR 99021

GMP Version Available: TB4423-GMP
GMP
Catalog # Availability Size / Price Qty
4423/10
4423/50
CHIR 99021 | CAS No. 252917-06-9 | GSK-3 Inhibitors
2 Images
Description: Highly selective GSK-3 inhibitor; acts as Wnt activator
Alternative Names: CHIR99021,CT99021

Chemical Name: 6-[[2-[[4-(2,4-Dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]-3-pyridinecarbonitrile

Purity: ≥98%

Product Details
Citations (412)
Supplemental Products
Reviews (2)

Biological Activity

CHIR 99021 is a potent and highly selective inhibitor of glycogen synthase kinase 3 (GSK-3) (IC50 values are 6.7 and 10 nM for GSK-3β and GSK-3α respectively). Acts as Wnt activator. Exhibits >500-fold selectivity for GSK-3 over closely related kinases; also displays >800-fold selectivity against 45 additional enzymes and receptors. In combination with the irreversible inhibitor of LSD1 (Cat. No. 3852), CHIR 99021 enables reprogramming of mouse embryonic fibroblasts, transduced by Oct4 and Klf4 only, into iPSCs. Enhances mouse and human ESC self-renewal when used in combination with PD 0325901 (Cat. No. 4192). It promotes differentiation of hPSCs in cardiomyocytes and allows formation of extended pluripotent stem (EPS) cells in combination with (S)-(+)-Dimethindene maleate (Cat. No. 1425), tetracycline antibiotic (Cat. No. 3268) and human leukemia inhibitory factor. Used in a protocol to generate brain organoids from human iPSCs. Also used in iPS cell differentiation protocol that produces CD34+ hematopoietic cells with multilineage engraftment ability.

CHIR 99021 synthesized to cGMP guidelines and CHIR 99021 in solution also available.

For more information on how to use CHIR 99021, see our protocols: Transdifferentiating Fibroblasts into Neurons, Culturing Transcriptomically Distinct Pluripotent mESCs, Converting Fibroblasts into Cardiomyocytes (9C Cocktail), Human iPSC-derived Myoblasts.

Scientific Data

Cardiomyocyte derived from human induced pluripotent stem cells (hiPSCs) using CHIR 99021. Cells cultured in Media supplemented with Activin A (Catalog # 338-AC, R&D Systems), N21-MAX Insulin Free (Catalog # AR010, R&D Systems), BMP-4 (Catalog # 314-BP, R&D Systems), bFGF (Catalog # 233-FB, R&D Systems) and DKK-1 (Catalog # 5439-DK, R&D Systems). Stained for F-Actin (Phalloidin, green), Troponin T (red, Human Troponin T (Cardiac) Antibody, Catalog # MAB1874, R&D Systems) and counterstained with DAPI (blue, Catalog # 5748, Tocris).

External Portal Information

Chemicalprobes.org is a portal that offers independent guidance on the selection and/or application of small molecules for research. The use of CHIR-99021 is reviewed on the chemical probes website.

Technical Data

M.Wt:
465.34
Formula:
C22H18Cl2N8
Solubility:
Soluble to 20 mM in DMSO
Purity:
≥98%
Storage:
Store at -20°C
CAS No:
252917-06-9

The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.

Background References

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    Xia Y, Sancho-Martinez I, Nivet E et al.
    Nat Protoc.
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  4. Efficient Derivation of Functional Human Airway Epithelium from Pluripotent Stem Cells via Temporal Regulation of Wnt Signaling
    KB McCauley, F Hawkins, M Serra, DC Thomas, A Jacob, DN Kotton
    Cell Stem Cell, 2017;0(0):.
  5. Enhanced Utilization of Induced Pluripotent Stem Cell-Derived Human Intestinal Organoids Using Microengineered Chips
    MJ Workman, JP Gleeson, EJ Troisi, HQ Estrada, SJ Kerns, CD Hinojosa, GA Hamilton, SR Targan, CN Svendsen, RJ Barrett
    Cell Mol Gastroenterol Hepatol, 2018;5(4):669-677.e2.
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    Orlova, Valeria, Drabsch, Yvette, Freund, Christia, Petrus-Reurer, Sandra, van den Hil, Francijn, Muenthaisong, Suchitra, Dijke, Peter Te, Mummery, Christin
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    Hill E, Ng T, Burton B, Oakley C, Malik K, Wraith D
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  12. Single-Cell Sequencing of iPSC-Dopamine Neurons Reconstructs Disease Progression and Identifies HDAC4 as a Regulator of Parkinson Cell Phenotypes
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  13. An Activating STAT3 Mutation Causes Neonatal Diabetes through Premature Induction of Pancreatic Differentiation
    J Saarimäki-, D Balboa, MA Russell, J Saarikettu, M Kinnunen, S Keskitalo, A Malhi, C Valensisi, C Andrus, S Eurola, H Grym, J Ustinov, K Wartiovaar, RD Hawkins, O Silvennoin, M Varjosalo, NG Morgan, T Otonkoski
    Cell Rep, 2017;19(2):281-294.
  14. The COPII cargo adapter SEC24C is essential for neuronal homeostasis
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    Nat Protoc, 2017;12(1):195-207.
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  18. Evaluating S. flexneri pathogenesis in the human enteroid model
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  20. Guided self-organization and cortical plate formation in human brain organoids
    MA Lancaster, NS Corsini, S Wolfinger, EH Gustafson, AW Phillips, TR Burkard, T Otani, FJ Livesey, JA Knoblich
    Nat. Biotechnol., 2017;0(0):.
  21. Renal Subcapsular Transplantation of PSC-Derived Kidney Organoids Induces Neo-vasculogenesis and Significant Glomerular and Tubular Maturation In�Vivo
    CW van den Be, L Ritsma, MC Avramut, LE Wiersma, BM van den Be, DG Leuning, E Lievers, M Koning, JM Vanslambro, AJ Koster, SE Howden, M Takasato, MH Little, TJ Rabelink
    Stem Cell Reports, 2018;0(0):.
  22. A Multiwell Cardiac ?GMEA Platform for Action Potential Recordings from Human iPSC-Derived Cardiomyocyte Constructs
    SL Edwards, V Zlochiver, DB Conrad, R Vaidyanath, AM Valiquette, R Joshi-Mukh
    Stem Cell Reports, 2018;0(0):.
  23. Prospective isolation of NKX2-1-expressing human lung progenitors derived from pluripotent stem cells
    F Hawkins, P Kramer, A Jacob, I Driver, DC Thomas, KB McCauley, N Skvir, AM Crane, AA Kurmann, AN Hollenberg, S Nguyen, BG Wong, AS Khalil, SX Huang, S Guttentag, JR Rock, JM Shannon, BR Davis, DN Kotton
    J. Clin. Invest., 2017;127(6):2277-2294.
  24. Comparative Analysis and Refinement of Human PSC-Derived Kidney Organoid Differentiation with Single-Cell Transcriptomics
    H Wu, K Uchimura, EL Donnelly, Y Kirita, SA Morris, BD Humphreys
    Cell Stem Cell, 2018;0(0):.
  25. Efficient generation of NKX6-1+ pancreatic progenitors from multiple human pluripotent stem cell lines.
    Nostro M, Sarangi F, Yang C, Holland A, Elefanty A, Stanley E, Greiner D, Keller G
    Stem Cell Reports, 2015;4(4):591-604.
  26. Lineage-Specific Chiral Biases of Human Embryonic Stem Cells during Differentiation
    KE Worley, AS Chin, LQ Wan
    Stem Cells Int, 2018;2018(0):1848605.
  27. Self-Renewing Trophoblast Organoids Recapitulate the Developmental Program of the Early Human Placenta
    S Haider, G Meinhardt, L Saleh, V Kunihs, M Gamperl, U Kaindl, A Ellinger, TR Burkard, C Fiala, J Pollheimer, S Mendjan, PA Latos, M Knöfler
    Stem Cell Reports, 2018;0(0):.
  28. A Regulatory Network Involving beta-Catenin, e-Cadherin, PI3k/Akt, and Slug Balances Self-Renewal and Differentiation of Human Pluripotent Stem Cells In Response to Wnt Signaling.
    Huang T, Li L, Moalim-Nour L, Jia D, Bai J, Yao Z, Bennett S, Figeys D, Wang L
    Stem Cells, 2015;33(5):1419-33.
  29. Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells.
    Orlova, Valeria, van den Hil, Francijn, Petrus-Reurer, Sandra, Drabsch, Yvette, Ten Dijke, Peter, Mummery, Christin
    Nat Protoc, 2014;9(6):1514-31.
  30. Derivation and characterization of putative craniofacial mesenchymal progenitor cells from human induced pluripotent stem cells
    M Jamal, SL Lewandowsk, ML Lawton, GT Huang, L Ikonomou
    Stem Cell Res, 2018;33(0):100-109.
  31. Nephron organoids derived from human pluripotent stem cells model kidney development and injury.
    Morizane R, Lam A, Freedman B, Kishi S, Valerius M, Bonventre J
    Nat Biotechnol, 2015;33(11):1193-200.
  32. Derivation and expansion of PAX7-positive muscle progenitors from human and mouse embryonic stem cells.
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    Stem Cell Reports, 2014;3(3):516-29.
  33. Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells
    F Sampazioti, MC de Brito, I Geti, A Bertero, NR Hannan, L Vallier
    Nat Protoc, 2017;12(4):814-827.
  34. 3D Modeling of Esophageal Development using Human PSC-Derived Basal Progenitors Reveals a Critical Role for Notch Signaling
    Y Zhang, Y Yang, M Jiang, SX Huang, W Zhang, D Al Alam, S Danopoulos, M Mori, YW Chen, R Balasubram, SM Chuva de S, C Serra, M Bialecka, E Kim, S Lin, ALR Toste de C, PN Riccio, WV Cardoso, X Zhang, HW Snoeck, J Que
    Cell Stem Cell, 2018;0(0):.
  35. Identification of direct negative crosstalk between the SLIT2 and Bone Morphogenetic Protein-Gremlin signaling pathways
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    J. Biol. Chem., 2018;0(0):.
  36. Heparin Promotes Cardiac Differentiation of Human Pluripotent Stem Cells in Chemically Defined Albumin-Free Medium, Enabling Consistent Manufacture of Cardiomyocytes

    Stem Cells Transl Med, 2016;0(0):.
  37. Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells
    N Tüysüz, L van Bloois, S van den Br, H Begthel, MM Verstegen, LJ Cruz, L Hui, LJ van der La, J de Jonge, R Vries, E Braakman, E Mastrobatt, JJ Cornelisse, H Clevers, D Ten Berge
    Nat Commun, 2017;8(0):14578.
  38. BMP and TGF-beta pathway mediators are critical upstream regulators of Wnt signaling during midbrain dopamine differentiation in human pluripotent stem cells.
    Cai J, Schleidt S, Pelta-Heller J, Hutchings D, Cannarsa G, Iacovitti L
    Dev Biol, 2013;376(1):62-73.
  39. Barrier properties and transcriptome expression in human iPSC-derived models of the blood-brain barrier
    L Delsing, P Dönnes, J Sánchez, M Clausen, D Voulgaris, A Falk, A Herland, G Brolén, H Zetterberg, R Hicks, J Synnergren
    Stem Cells, 2018;0(0):.
  40. Differentiation of human limbal-derived induced pluripotent stem cells into limbal-like epithelium.
    Sareen D, Saghizadeh M, Ornelas L, Winkler M, Narwani K, Sahabian A, Funari V, Tang J, Spurka L, Punj V, Maguen E, Rabinowitz Y, Svendsen C, Liubimov A
    Stem Cells Transl Med, 2014;3(9):1002-12.
  41. Generation of Mouse and Human Organoid-Forming Intestinal Progenitor Cells by Direct Lineage Reprogramming
    S Miura, A Suzuki
    Cell Stem Cell, 2017;0(0):.
  42. Phenotypic and Functional Characterization of Peripheral Sensory Neurons derived from Human Embryonic Stem Cells
    AJ Alshawaf, S Viventi, W Qiu, G D'Abaco, B Nayagam, M Erlichster, G Chana, I Everall, J Ivanusic, E Skafidas, M Dottori
    Sci Rep, 2018;8(1):603.
  43. AKT kinase activity is required for Li to modulate mood-related behaviors in mice.
    Pan et al.
    Neuropsychopharmacology, 2011;36:1397
  44. Pleiotropy of glycogen synthase kinase-3 inhibition by CHIR99021 promotes self-renewal of embyronic stem cells from refractory mouse strains.
    Ye et al.
    PLoS One, 2012;7:e35892
  45. Generation of human-induced pluripotent stem cells in the absence of exogenous Sox2.
    Li et al.
    Stem Cells, 2009;27:2992
  46. Derivation of pluripotent stem cells with in vivo embryonic and extraembryonic potency.
    Yang et al.
    Cell., 2017;169:243
  47. Controlling Epithelial Polarity: A Human Enteroid Model for Host-Pathogen Interactions
    JY Co, M Margalef-C, X Li, AT Mah, CJ Kuo, DM Monack, MR Amieva
    Cell Rep, 2019;26(9):2509-2520.e4.
  48. Enhanced Utilization of Induced Pluripotent Stem Cell-Derived Human Intestinal Organoids Using Microengineered Chips.
    Michael J Workman, John P Gleeson, Elissa J Troisi, Hannah Q Estrada, S Jordan Kerns, Christopher D Hinojosa, Geraldine A Hamilton, Stephan R Targan, Clive N Svendsen, Robert J Barrett
    Cellular and Molecular Gastroenterology and Hepatology, 2018;0(0):2352-345X.
  49. In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection.
    Bartfeld et al.
    Gastroenterology, 2015;148:126
  50. SnapShot: Growing Organoids from Stem Cells.
    Sato et al.
    Cell, 2015;161:1700
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    Mahe et al.
    Curr.Protoc. Mouse Biol., 2014;3:217
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    Dye et al.
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    Wimmer et al.
    Nature, 2019;565:505
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    Noor et al.
    Adv.Sci.(Weinh), 2019;6:1900344
  55. Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells.
    Hoang et al.
    Nat.Protoc., 2018;13:723
  56. Directed differentiation of human induced pluripotent stem cells into mature stratified bladder urothelium
    K Suzuki, M Koyanagi-A, K Uehara, N Hinata, M Fujisawa, T Aoi
    Sci Rep, 2019;9(1):10506.
  57. Chronic aerobic exercise improves insulin sensitivity and modulates Nrf2 and NF?kappaB/IkappaBalpha pathways in the skeletal muscle of rats fed with a high fat diet
    Q Yu, Z Xia, EC Liong, GL Tipoe
    Mol Med Rep, 2019;20(6):4963-4972.
  58. Cyclin G2 regulates canonical Wnt signalling via interaction with Dapper1 to attenuate tubulointerstitial fibrosis in diabetic nephropathy
    C Zhao, J Gao, S Li, Q Liu, X Hou, X Xing, D Wang, M Sun, S Wang, Y Luo
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    Mol. Ther., 2020;28(1):129-141.
  60. Multipotent RAG1+ progenitors emerge directly from haemogenic endothelium in human pluripotent stem cell-derived haematopoietic organoids
    A Motazedian, FF Bruveris, SV Kumar, JV Schiesser, T Chen, ES Ng, AP Chidgey, CA Wells, AG Elefanty, EG Stanley
    Nat. Cell Biol., 2020;22(1):60-73.
  61. Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes
    S Selvaraj, R Mondragon-, B Xu, A Magli, H Kim, J Lainé, J Kiley, H Mckee, F Rinaldi, J Aho, N Tabti, W Shen, RC Perlingeir
    Elife, 2019;8(0):.
  62. ER stress reliever enhances functionalities of in vitro cultured hepatocytes
    JS Kim, SI Hwang, JL Ryu, HS Hong, JM Lee, SM Lee, X Jin, C Han, JH Kim, J Han, MR Lee, DH Woo
    Stem Cell Res, 2020;43(0):101732.
  63. Automated feature extraction from large cardiac electrophysiological data sets
    J Jurkiewicz, S Kroboth, V Zlochiver, P Hinow
    Journal of electrocardiology, 2021;65(0):157-162.
  64. Hyperglycemia Altered DNA Methylation Status and Impaired Pancreatic Differentiation from Embryonic Stem Cells
    ACH Chen, W Huang, SW Fong, C Chan, KC Lee, WSB Yeung, YL Lee
    International Journal of Molecular Sciences, 2021;22(19):.
  65. Tissue context determines the penetrance of regulatory DNA variation
    JM Halow, R Byron, MS Hogan, R Ordoñez, M Groudine, MA Bender, JA Stamatoyan, MT Maurano
    Nature Communications, 2021;12(1):2850.
  66. Auto/paracrine factors and early Wnt inhibition promote cardiomyocyte differentiation from human induced pluripotent stem cells at initial low cell density
    MNT Le, M Takahi, K Ohnuma
    Scientific Reports, 2021;11(1):21426.
  67. High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids
    S Xia, Z Bozóky, M Di Paola, O Laselva, S Ahmadi, JX Jiang, AL Pitstick, C Jiang, D Rotin, CN Mayhew, NL Jones, CE Bear
    Cells, 2021;10(12):.
  68. Human induced pluripotent stem cells integrate, create synapses and extend long axons after spinal cord injury
    NS Lavoie, V Truong, D Malone, T Pengo, N Patil, JR Dutton, AM Parr
    Journal of Cellular and Molecular Medicine, 2022;0(0):.
  69. Development of High affinity Nanobodies Specific for NaV1.4 and NaV1.5 Voltage-Gated Sodium Channel Isoforms
    L Srinivasan, V Alzogaray, D Selvakumar, S Nathan, JB Yoder, KM Wright, S Klinke, JN Nwafor, MS Labanda, FA Goldbaum, A Schön, E Freire, GF Tomaselli, LM Amzel, M Ben-Johny, SB Gabelli
    The Journal of Biological Chemistry, 2022;0(0):101763.

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Citations for CHIR 99021

The citations listed below are publications that use Tocris products. Selected citations for CHIR 99021 include:

412 Citations: Showing 1 - 10

  1. Metformin decelerates aging clock in male monkeys.
    Authors: Yang Et al.
    Cell  2024;187:1
  2. Long-term engrafting multilineage hematopoietic cells differentiated from human induced pluripotent stem cells.
    Authors: Ng Et al.
    Nat.Biotechnol.  2024;
  3. In vitro reconstitution of epigenetic reprogramming in the human germ line.
    Authors: Murase Et al.
    Nature  2024;
  4. Pluripotent stem cell-derived committed cardiac progenitors remuscularize damaged ischemic hearts and improve their function in pigs.
    Authors: Jing Et al.
    NPJ Regen Med  2023;8:26
  5. Systematic discovery of transcription factors that improve hPSC-derived cardiomyocyte maturation via temporal analysis of bioengineered cardiac tissues.
    Authors: Aditya Et al.
    APL Bioeng  2023;7:026109
  6. Spatial multiomics map of trophoblast development in early pregnancy.
    Authors: Qian Et al.
    Nature  2023;616:143-151
  7. Engineered cardiac tissue model of restrictive cardiomyopathy for drug discovery.
    Authors: Yulia V Et al.
    Cell Rep Med  2023;4:100976
  8. A Robust Pipeline for the Multi-Stage Accelerated Differentiation of Functional 3D Cortical Organoids from Human Pluripotent Stem Cells.
    Authors: Mustafa Et al.
    Curr Protoc  2023;3:e641
  9. Quantitative chemometric phenotyping of three-dimensional liver organoids by Raman spectral imaging.
    Authors: Stefan Et al.
    Cell Rep Methods  2023;3:100440
  10. Genetic dissection of the pluripotent proteome through multi-omics data integration.
    Authors: Steven P Et al.
    Cell Genom  2023;3:100283
  11. Automated high-content imaging in iPSC-derived neuronal progenitors.
    Authors: Serena Et al.
    SLAS Discov  2023;28:42-51
  12. Generation of dorsoventral human spinal cord organoids via functionalizing composite scaffold for drug testing.
    Authors: Bo Et al.
    iScience  2023;26:105898
  13. Expansion of ventral foregut is linked to changes in the enhancer landscape for organ-specific differentiation.
    Authors: Sara Et al.
    Nat Cell Biol  2023;25:481-492
  14. Long-Term SMN- and Ncald-ASO Combinatorial Therapy in SMA Mice and NCALD-ASO Treatment in hiPSC-Derived Motor Neurons Show Protective Effects.
    Authors: C Frank Et al.
    Int J Mol Sci  2023;24
  15. Airway and Lung Organoids from Human-Induced Pluripotent Stem Cells Can Be Used to Assess CFTR Conductance.
    Authors: Dmitry Et al.
    Int J Mol Sci  2023;24
  16. Pump-less, recirculating organ-on-a-chip (rOoC) platform.
    Authors: Stefan Et al.
    Lab Chip  2023;23:591-608
  17. FUS-ALS hiPSC-derived astrocytes impair human motor units through both gain-of-toxicity and loss-of-support mechanisms.
    Authors: Ludo Et al.
    Mol Neurodegener  2023;18:5
  18. Bioengineering human intestinal mucosal grafts using patient-derived organoids, fibroblasts and scaffolds.
    Authors: Laween Et al.
    Nat Protoc  2023;18:108-135
  19. An in vivo neuroimmune organoid model to study human microglia phenotypes.
    Authors: Schafer Et al.
    Cell  2023;186:2111
  20. A pesticide and iPSC dopaminergic neuron screen identifies and classifies Parkinson-relevant pesticides.
    Authors: Tim Et al.
    Nat Commun  2023;14:2803
  21. Establishing Bipotential Human Lung Organoid Culture System and Differentiation to Generate Mature Alveolar and Airway Organoids.
    Authors: Jie Et al.
    Bio Protoc  2023;13:e4657
  22. Axon guidance genes modulate neurotoxicity of ALS-associated UBQLN2.
    Authors: Mark Et al.
    Elife  2023;12
  23. Establishment of Skeletal Myogenic Progenitors from Non-Human Primate Induced Pluripotent Stem Cells.
    Authors: Alessandro Et al.
    Cells  2023;12
  24. Recapitulating porcine cardiac development in vitro: from expanded potential stem cell to embryo culture models.
    Authors: Alessandra Et al.
    Front Cell Dev Biol  2023;11:1111684
  25. A distal lung organoid model to study interstitial lung disease, viral infection and human lung development.
    Authors: Nadine Et al.
    Nat Protoc  2023;
  26. In vivo development of immune tissue in human intestinal organoids transplanted into humanized mice
    Authors: Bouffi Et al.
    Nat.Biotechnol.  2023;
  27. Ti3 C2 Tx MXene Composite 3D Hydrogel Potentiates mTOR Signaling to Promote the Generation of Functional Hair Cells in Cochlea Organoids.
    Authors: Chen Et al.
    Adv Sci (Weinh)  2022;9:e2203557
  28. Transcription Factor Hb9 Is Expressed in Glial Cell Lineages in the Developing Mouse Spinal Cord.
    Authors: Diego Et al.
    Eneuro  2022;9
  29. Engineering a living cardiac pump on a chip using high-precision fabrication.
    Authors: Kamil Et al.
    Sci Adv  2022;8:eabm3791
  30. Transplanted organoids empower human preclinical assessment of drug candidate for the clinic.
    Authors: Michael Et al.
    Sci Adv  2022;8:eabj5633
  31. Reengineering Ponatinib to Minimize Cardiovascular Toxicity.
    Authors: Wenqi Et al.
    Cancer Res  2022;82:2777-2791
  32. JAK inhibitor blocks COVID-19 cytokine-induced JAK/STAT/APOL1 signaling in glomerular cells and podocytopathy in human kidney organoids.
    Authors: Guojie Et al.
    JCI Insight  2022;7
  33. The antimicrobial peptide S100A8/A9 produced by airway epithelium functions as a potent and direct regulator of macrophage phenotype and function.
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iPSC differentiation
By Ricardo Malvicini on 09/26/2021
Species: Human

Used for the differentiation of Human IPSc to cardiomyocites (concentrarion of 5uM)


cells treatment
By Anonymous on 12/04/2017

Treating cells


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