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Submit ReviewY-27632 dihydrochloride
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Y-27632 dihydrochloride
Description: Selective ROCK inhibitor; inhibits ROCK1 (p160 ROCK) and ROCK2; stem cell freezing (cryopreservation) media component
Alternative Names: y27632,Y-27632 2HCl,ROCK inhibitor,ROCK1 inhibitor
Chemical Name: trans-4-[(1R)-1-Aminoethyl]-N-4-pyridinylcyclohexanecarboxamide dihydrochloride
Purity: ≥98% (HPLC)
Biological Activity for Y-27632 dihydrochloride
Y-27632 dihydrochloride is a selective ROCK inhibitor (Ki values are 0.14-0.22, 0.3, 25, 26 and > 250 μM for ROCK1 (p160 ROCK), ROCK2, PKA, PKC and MLCK respectively). Also inhibits PRK2 (IC50 = 600 nM). Increases survival rate of human embryonic stem (hES) cells and iPSC undergoing cryopreservation. Used in combination with CHIR 99021, RepSox, Forskolin, SP 600125, Go 6983 and VPA to reprogram fibroblasts to mature neurons. Also optimizes naive human pluripotent stem cell growth and viability following naive cell derivation from primed ESCs and iPSCs using naive human stem cell medium (NHSM). Used as component of growth media for urothelial organoids. Also used in a protocol to generate brain organoids from human iPSCs. Cell-permeable, orally bioavailable and active in vivo.
Y-27632 in solution and Y-27632 synthesized to cGMP guidelines also available.
For more information about how Y-27632 dihydrochloride may be used, see our protocols: Transdifferentiating Fibroblasts into Neurons, Converting Fibroblasts into Cardiomyocytes (9C Cocktail), Accelerated Induction of Cortical Neurons from hiPSCs, Human iPSC-derived Myoblasts.
Scientific Data
Application of Y-27632 in MESP1+ Mesoderm. ROCK inhibition by Y-27632 (1 μM) promotes skeletal myogenic differentiation of MESP1+ mesoderm, as shown by immunostaining for Myosin Heavy Chain+ (Green) and Myogenin (Red), and counterstaining with DAPI (Catalog # 5748; Blue, Tocris). Data courtesy of Dr Sunny Chan and Dr Michael Kyba, University of Minnesota.
Compound Libraries for Y-27632 dihydrochloride
Y-27632 dihydrochloride is also offered as part of the Tocriscreen 2.0 Max, Tocriscreen Kinase Inhibitor Library and Tocriscreen Stem Cell Library. Find out more about compound libraries available from Tocris.
Technical Data for Y-27632 dihydrochloride
| M. Wt | 320.26 |
| Formula | C14H21N3O.2HCl |
| Storage | Desiccate at RT |
| Purity | ≥98% (HPLC) |
| CAS Number | 129830-38-2 |
| PubChem ID | 14801906 |
| InChI Key | IDDDVXIUIXWAGJ-LJDSMOQUSA-N |
| Smiles | N[C@H](C)[C@@]1([H])CC[C@H](C(NC2=CC=NC=C2)=O)CC1.Cl.Cl |
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.
Solubility Data for Y-27632 dihydrochloride
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 32.03 | 100 | |
| water | 32.03 | 100 |
Preparing Stock Solutions for Y-27632 dihydrochloride
The following data is based on the product molecular weight 320.26. Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
|---|---|---|---|
| 1 mM | 3.12 mL | 15.61 mL | 31.22 mL |
| 5 mM | 0.62 mL | 3.12 mL | 6.24 mL |
| 10 mM | 0.31 mL | 1.56 mL | 3.12 mL |
| 50 mM | 0.06 mL | 0.31 mL | 0.62 mL |
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Product Datasheets for Y-27632 dihydrochloride
References for Y-27632 dihydrochloride
References are publications that support the biological activity of the product.
Narumiya et al (2000) Use and properties of ROCK-specific inhibitor Y-27632. Methods Enzymol. 325 273 PMID: 11036610
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Ishizaki et al (2000) Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol.Pharmacol. 57 976 PMID: 10779382
Hu et al (2015) Direct conversion of normal and Alzheimer's Disease human fibroblasts into neuronal cells by small molecules. Cell Stem Cell 17 204 PMID: 26253202
Chan et al et al (2016) Development of bipotent cardiac/skeletal myogenic progenitors from MESP1+ mesoderm. Stem Cell Rep. 6 26 PMID: 26771351
Santos et al (2019) Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity. Nat.Commun. 10 4407 PMID: 31562298
Schafer (2023) An in vivo neuroimmune organoid model to study human microglia phenotypes. Cell 186 2111 PMID: 37172564
If you know of a relevant reference for Y-27632 dihydrochloride, please let us know.
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252 Citations for Y-27632 dihydrochloride
Citations are publications that use Tocris products. Selected citations for Y-27632 dihydrochloride include:
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Hubaud et al (2017) Excitable Dynamics and Yap-Dependent Mechanical Cues Drive the Segmentation Clock. Cell 171 668 PMID: 28942924
Bershteyn et al (2017) Human iPSC-Derived Cerebral Organoids Model Cellular Features of Lissencephaly and Reveal Prolonged Mitosis of Outer Radial Glia. Cell Stem Cell 20 435 PMID: 28111201
Sloan et al (2017) Human Astrocyte Maturation Captured in 3D Cerebral Cortical Spheroids Derived from Pluripotent Stem Cells. Neuron 95 779 PMID: 28817799
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Shukla et al (2017) ASXL3 Is a Novel Pluripotency Factor in Human Respiratory Epithelial Cells and a Potential Therapeutic Target in Small Cell Lung Cancer. Cancer Res 77 6267 PMID: 28935813
Bao et al (2017) Directed differentiation and long-term maintenance of epicardial cells derived from human pluripotent stem cells under fully defined conditions. Nat Protoc 12 1890 PMID: 28817124
Vermilyea et al (2017) Induced Pluripotent Stem Cell-Derived Dopaminergic Neurons from Adult Common Marmoset Fibroblasts. Stem Cells Dev 26 1225 PMID: 28635509
McCauley et al (2017) Efficient Derivation of Functional Human Airway Epithelium from Pluripotent Stem Cells via Temporal Regulation of Wnt Signaling. Cell Stem Cell 20 844 PMID: 28366587
Tischfield et al (2017) Atypical PKC and Notch Inhibition Differentially Modulate Cortical Interneuron Subclass Fate from Embryonic Stem Cells. Stem Cell Reports 8 1135 PMID: 28416285
Yang et al (2017) Derivation of Pluripotent Stem Cells with In Vivo Embryonic and Extraembryonic Potency. Cell 169 243 PMID: 28388409
Morizane and Bonventre (2017) Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells. Nat Protoc 12 195 PMID: 28005067
Kinarivala et al (2017) Discovery of Aromatic Carbamates that Confer Neuroprotective Activity by Enhancing Autophagy and Inducing the Anti-Apoptotic Protein B-Cell Lymphoma 2 (Bcl-2). J Med Chem 60 9739 PMID: 29110485
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Aguisanda et al (2017) Neural stem cells for disease modeling of Wolman disease and evaluation of therapeutics. Orphanet J Rare Dis 12 120 PMID: 28659158
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Sharma et al (2019) Exosomes regulate neurogenesis and circuit assembly. Proc Natl Acad Sci U S A 116 16086 PMID: 31320591
Vattulainen et al (2019) Modulation of Wnt/BMP pathways during corneal differentiation of hPSC maintains ABCG2-positive LSC population that demonstrates increased regenerative potential. Stem Cell Res Ther 10 236 PMID: 31383008
Wang et al (2019) Protective effects of oxymatrine against DSS-induced acute intestinal inflammation in mice via blocking the RhoA/ROCK signaling pathway. Biosci Rep 39 PMID: 31262973
Hladik et al (2019) Long-term culture of mesenchymal stem cells impairs ATM-dependent recognition of DNA breaks and increases genetic instability. Stem Cell Res Ther 10 218 PMID: 31358047
Wilke et al (2019) A Stem-Cell-Derived Platform Enables Complete Cryptosporidium Development In Vitro and Genetic Tractability. Cell Host Microbe 26 123 PMID: 31231046
Rehbach et al (2019) Multiparametric rapid screening of neuronal process pathology for drug target identification in HSP patient-specific neurons. Sci Rep 9 9615 PMID: 31270336
Lee et al (2019) Entosis Controls a Developmental Cell Clearance in C. elegans. Cell Rep 26 3212 PMID: 30893595
Montel-Hagen et al (2019) Organoid-Induced Differentiation of Conventional T Cells from Human Pluripotent Stem Cells. Cell Stem Cell 24 376 PMID: 30661959
Li et al (2019) A Non-canonical Pathway with Potential for Safer Modulation of Transforming Growth Factor-β1 in Steroid-Resistant Airway Diseases. iScience 12 232 PMID: 30711747
Maguire et al (2019) Highly Efficient CRISPR-Cas9-Mediated Genome Editing in Human Pluripotent Stem Cells. Curr Protoc Stem Cell Biol 48 e64 PMID: 30358158
Frith and Tsakiridis (2019) Efficient Generation of Trunk Neural Crest and Sympathetic Neurons from Human Pluripotent Stem Cells Via a Neuromesodermal Axial Progenitor Intermediate. Curr Protoc Stem Cell Biol e81 PMID: 30688409
Benedicto (2017) Concerted regulation of retinal pigment epithelium basement membrane and barrier function by angiocrine factors. Nat Commun 8 15374 PMID: 28524846
Xi (2017) In Vivo Human Somitogenesis Guides Somite Development from hPSCs. Cell Rep 18 1573 PMID: 28178531
Hall (2017) Mesoderm-specific Stat3 deletion affects expression of Sox9 yielding Sox9-dependent phenotypes. PLoS Genet 13 e1006610 PMID: 28166224
Kotton et al (2017) Differentiation of human pluripotent stem cells into functional lung alveolar epithelial cells. Cell Stem Cell. 21 472 PMID: 28965766
Tchieu et al (2017) A modular platform for differentiation of human PSCs into all major ectodermal lineages. Cell Stem Cell. 31 399 PMID: 28886367
Shi (2017) The novel cannabinoid receptor GPR55 mediates anxiolytic-like effects in the medial orbital cortex of mice with acute stress. Mol Brain 10 38 PMID: 28800762
Rempel et al (2015) A transcriptome-based classifier to identify developmental toxicants by stem cell testing: design, validation and optimization for histone deacetylase inhibitors. Arch Toxicol 89 1599 PMID: 26272509
Loubet et al (2012) Neuritogenesis: the prion protein controls β1 integrin signaling activity. FASEB J 26 678 PMID: 22038049
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Prahalad et al (2004) Regulation of MDCK cell-substratum adhesion by RhoA and myosin light chain kinase after ATP depletion. Am J Physiol Cell Physiol 286 C693 PMID: 14644769
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Wibberley et al (2003) Expression and functional role of Rho-kinase in rat urinary bladder smooth muscle. Br J Pharmacol 138 757 PMID: 12642376
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Ng-Blichfeldt et al (2019) TGF-β activation impairs fibroblast ability to support adult lung epithelial progenitor cell organoid formation. Am J Physiol Lung Cell Mol Physiol PMID: 30969812
Mair et al (2019) Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies. Cell Rep 27 599 PMID: 30970261
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Bertero et al (2019) Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory. Nat Commun 10 1538 PMID: 30948719
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Hackland et al (2019) FGF Modulates the Axial Identity of Trunk hPSC-Derived Neural Crest but Not the Cranial-Trunk Decision. Stem Cell Reports 12 920 PMID: 31091435
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Reviews for Y-27632 dihydrochloride
Average Rating: 5 (Based on 14 Reviews.)
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Y-27632.
By
Chi Ngo on 11/25/2024
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: human iPSC
I used the Y-27632 compound to help with iPSC's viability after thaw.
Consistent performance.
By
Ted Mason on 08/22/2023
Assay Type: In Vitro
Species: Human
ROCKi has consistently performed in iPSC cell culture work. Well packaged vials are resuspended and aliquoted for -20 storage. Cells passaged or fed with ROCKi media have shown good viability. This is a regular purchase for the labs at a reasonable price.Images @10x magnification show limited differentiation of iPSCs after 3 days in culture.
Excellent product.
By
Jack Schnell on 03/15/2023
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: iPSCs
We add this product at a concentration of 10 uM to promote cell survival in in vitro iPSC cultures.
Easy to manage and works as expected
ROCK Inhibitor.
By
Anonymous on 09/07/2020
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: Organoids
To generate human intestinal organoids
test.
By
Veselina Petrova on 08/28/2020
Assay Type: In Vitro
Species: Human
I used this product on freshly split and plate human iPSC cells to help cell survival and adherence. I used the product at 10 mM concentration which I diluted in 1:1000 in culture media and left the cells at 37C, 5% CO2 for 24 hours before replacing with fresh culture media.
For hPSC culture.
By
Anonymous on 06/30/2020
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: Human PSC
For hPSC culture, final concentration 10mM. Reconstitute 10 mg of Y27632 in 3.0792 ml of PBS. Make 100µl aliquots and keep them at −20 °C for up to 6 months.
Worked great and at a great price.
By
Tres Davis on 05/08/2020
Assay Type: Ex Vivo
Species: Mouse
HPLC
Neural Induction Conditions.
By
Anonymous on 03/22/2020
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: H9
ROCK inhibitor Y-27632 (10 mM)
Great for passaging stem cells..
By
Samiha Shaikh on 01/31/2020
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: iPSC
Great for passaging huiPSCs with dissociation into single cells.
Great for single iPS cell survival.
By
Anonymous on 09/13/2019
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: iPSC
Used during single cell passage for CRISPR clone isolation, works well and no difference found between multiple batches. Cells show characteristic 'spiky' morphology upon addition to the medium.
I use at 1:1000 and store at -20C. I remove the compound from medium 24 hours after plating.
For iPSC culture.
By
Pinyuan Tian on 08/07/2018
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: iPSC
For iPSC culture, final concentration 10mM. Reconstitute 10 mg of Y27632 in 3.0792 ml of PBS.
Make 100µl aliquots and keep them at −20 °C for up to 6 months.
ROCK Inhibitor to improve cartilage integrity.
By
Anonymous on 03/26/2018
Assay Type: In Vitro
Species: Other
Cell Line/Tissue: chondrocyte
I dissolved in DMSO
I prepare 20mg of the product in 5.88mL H2O and I use 1uL/mL of the stock in my cell culture after the cell splitting.
By
Anonymous on 02/01/2018
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: human foreskin fibroblasts-derived-IPS
I used it in two human IPS cell lines, 5uM concentration the first 24 h after the splitting. The use of this reagent improves substantially the cell survival.
Keep the stock frozen at -20°C. I use it freshly added to the medium when splitting the cells.
ROCK Inhibitor to improve cell survival.
By
Hudson DE OLIVEIRA BEZERRA on 01/05/2018
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: H1 (W01)
My team uses this product to improve human embryonic stem cells survival in single cell passage methods in siRNA and CRISPR experiments. In general, we use the concentration of 10 uM. The products work very well, better than others available in the market.
Protocols for Y-27632 dihydrochloride
The following protocols feature additional information for the use of Y-27632 dihydrochloride (Cat. No. 1254).
- Transdifferentiating Fibroblasts into Neurons
- Converting Fibroblasts into Cardiomyocytes (9C Cocktail)
- Accelerated Induction of Cortical Neurons from hiPSCs
- Human iPSC-derived Myoblasts
- Generation and Propagation of EPS cells (LCDM Cocktail)
- Cultivating Cerebral Organoids
- Generating Vascular Organoids
- 3D Culture of Lung Alveolar Cells
- Generating Midbrain Dopaminergic Neurons from hPSCs
Literature in this Area
Tocris offers the following scientific literature in this area 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.
Stem Cells Scientific Review
Written by Kirsty E. Clarke, Victoria B. Christie, Andy Whiting and Stefan A. Przyborski, this review provides an overview of the use of small molecules in the control of stem cell growth and differentiation. Key signaling pathways are highlighted, and the regulation of ES cell self-renewal and somatic cell reprogramming is discussed. Compounds available from Tocris are listed.
Stem Cell Workflow Poster
Stem cells have potential as a source of cells and tissues for research and treatment of disease. This poster summarizes some key protocols demonstrating the use of small molecules across the stem cell workflow, from reprogramming, through self-renewal, storage and differentiation to verification. Advantages of using small molecules are also highlighted.
Stem Cells Poster
Written by Rebecca Quelch and Stefan Przyborski from Durham University (UK), this poster describes the isolation of pluripotent stem cells, their maintenance in culture, differentiation, and the generation and potential uses of organoids.