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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 | |||
| water | 32.03 | 100 | |
| DMSO | 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.
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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
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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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Lynch et al (2018) Submucosal Gland Myoepithelial Cells Are Reserve Stem Cells That Can Regenerate Mouse Tracheal Epithelium. Cell Stem Cell 22 653 PMID: 29656941
Gupta et al (2018) CRISPR/Cas9-based Targeted Genome Editing for the Development of Monogenic Diseases Models with Human Pluripotent Stem Cells. Curr Protoc Stem Cell Biol 45 e50 PMID: 30040245
McCauley et al (2018) Derivation of Epithelial-Only Airway Organoids from Human Pluripotent Stem Cells. Curr Protoc Stem Cell Biol 45 e51 PMID: 30040246
Asumda et al (2018) Differentiation of hepatocyte-like cells from human pluripotent stem cells using small molecules. Differentiation 101 16 PMID: 29626713
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Mulay et al (2016) An in vitro model of murine middle ear epithelium. Dis Model Mech 9 1405 PMID: 27660200
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Stebbins et al (2016) Differentiation and characterization of human pluripotent stem cell-derived brain microvascular endothelial cells. Methods 101 93 PMID: 26518252
Zeltner et al (2016) Capturing the biology of disease severity in a PSC-based model of familial dysautonomia. Nat Med 22 1421 PMID: 27841875
Tidball et al (2016) Genomic Instability Associated with p53 Knockdown in the Generation of Huntington's Disease Human Induced Pluripotent Stem Cells. PLoS One 11 e0150372 PMID: 26982737
Kellner et al (2016) Nogo-A controls structural plasticity at dendritic spines by rapidly modulating actin dynamics. Hippocampus 6 816 PMID: 26748478
Chal et al (2016) Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro. Nat.Protoc. 11 1833 PMID: 27583644
Borgs et al (2016) DArgic neurons differentiating from LRRK2 G2019S induced pluripotent stem cells show early neuritic branching defects. Scientific Reports 6 33377 PMID: 27640816
Pattabiraman et al (2015) RhoA GTPase-induced ocular hypertension in a rodent model is associated with increased fibrogenic activity in the trabecular meshwork. Autophagy 185 496 PMID: 25499974
Lachmann et al (2015) Large-scale hematopoietic differentiation of human induced pluripotent stem cells provides granulocytes or macrophages for cell replacement therapies. Nat Commun 4 282 PMID: 25680479
Abraham et al (2015) A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis. Stem Cell Reports 6 7286 PMID: 26129894
Arbab et al (2015) Cloning-free CRISPR. Cell 5 908 PMID: 26527385
Ruprecht et al (2015) Cortical contractility triggers a stochastic switch to fast amoeboid cell motility. Int J Mol Sci 160 673 PMID: 25679761
Vergara et al (2015) Transgenic plants as low-cost platform for chemotherapeutic drugs screening. J Biol Chem 16 2174 PMID: 25608652
Lee et al (2015) Odontogenic Ameloblast-associated Protein (ODAM) Mediates Junctional Epithelium Attachment to Teeth via Integrin-ODAM-Rho Guanine Nucleotide Exchange Factor 5 (ARHGEF5)-RhoA Signaling. Toxicol Sci 290 14740 PMID: 25911094
Tyson et al (2015) Duration of culture and sonic hedgehog signaling differentially specify PV versus SST cortical interneuron fates from embryonic stem cells. Stem Cells 142 1267 PMID: 25804737
Balcioglu et al (2015) The integrin expression profile modulates orientation and dynamics of force transmission at cell-matrix adhesions. Nat Biotechnol 128 1316 PMID: 25663698
Morizane et al (2015) Nephron organoids derived from human pluripotent stem cells model kidney development and injury. Nat Commun 33 1193 PMID: 26458176
Chaudhuri et al (2015) Substrate stress relaxation regulates cell spreading. Brain 6 6364 PMID: 25695512
Iovino et al (2015) Early maturation and distinct tau pathology in induced pluripotent stem cell-derived neurons from patients with MAPT mutations. Development 138 3345 PMID: 26220942
Jönsson et al (2015) Comprehensive analysis of microRNA expression in regionalized human neural progenitor cells reveals microRNA-10 as a caudalizing factor. Int J Nanomedicine 142 3166 PMID: 26395143
Tai et al (2015) SimV. enhances Rho/actin/cell rigidity pathway contributing to mesenchymal stem cells' osteogenic differentiation. Nat Commun 10 5881 PMID: 26451103
Sakaguchi et al (2015) Generation of functional hippocampal neurons from self-organizing human embryonic stem cell-derived dorsomedial telencephalic tissue. Mol Vis 6 8896 PMID: 26573335
Su et al (2015) Phenotypes of trypsin- and Col. -prepared bovine corneal endothelial cells in the presence of a selective Rho kinase inhibitor, Y-27632. PLoS Comput Biol 21 633 PMID: 26097378
Ziv et al (2015) Quantitative Live Imaging of Human Embryonic Stem Cell Derived Neural Rosettes Reveals Structure-Function Dynamics Coupled to Cortical Development. Stem Cell Reports 11 e1004453 PMID: 26473351
Fernandes et al (2015) Comparison of Human Embryonic Stem Cell-Derived Cardiomyocytes, Cardiovascular Progenitors, and Bone Marrow Mononuclear Cells for Cardiac Repair. Bone Res NULL PMID: 26607951
Maharam et al (2015) Rho/Rock signal transduction pathway is required for MSC tenogenic differentiation. Stem Cell Res Ther 3 15015 PMID: 26509098
Luzzani et al (2015) A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement. Int J Cancer 6 6 PMID: 25582222
Choi et al (2015) SDF-1α stiffens myeloma bone marrow mesenchymal stromal cells through the activation of RhoA-ROCK-Myosin II. Stem Cell Reports 136 E219 PMID: 25137150
Höfner et al (2015) Defined conditions for the isolation and expansion of basal prostate progenitor cells of mouse and human origin. PLoS One 4 503 PMID: 25702639
Härmä et al (2015) Optimization of Invasion-Specific Effects of Betulin Derivatives on Prostate Cancer Cells through Lead Development. Stem Cell Reports 10 e0126111 PMID: 25965345
Gomez et al (2010) Tissue geometry patterns epithelial-mesenchymal transition via intercellular mechanotransduction. ACS Chem Neurosci 110 44 PMID: 20336666
Cao et al (2010) Cortical actin dynamics facilitate early-stage centrosome separation. J Pharmacol Exp Ther 20 770 PMID: 20409712
Kim et al (2010) PDGF signaling is required for epicardial function and blood vessel formation in regenerating zebrafish hearts. FEBS Lett 107 17206 PMID: 20858732
Surendran et al (2022) An improved protocol for generation and characterization of human-induced pluripotent stem cell-derived retinal pigment epithelium cells STAR Protoc 3 101803 PMID: 36386870
Wong et al (2018) α-Tubulin Acetyltransferase Is a Novel Target Mediating Neurite Growth Inhibitory Effects of Chondroitin Sulfate Proteoglycans and Myelin-Associated Glycoprotein. Eneuro 5 PMID: 29497702
Ho et al (2018) Lysophosphatidic Acid Induces ECM Production via Activation of the Mechanosensitive YAP/TAZ Transcriptional Pathway in Trabecular Meshwork Cells. Invest Ophthalmol Vis Sci 59 1969 PMID: 29677358
Sima et al (2018) Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses. Orphanet J Rare Dis 13 54 PMID: 29631617
Medelnik et al (2018) Signaling-Dependent Control of Apical Membrane Size and Self-Renewal in Rosette-Stage Human Neuroepithelial Stem Cells. Stem Cell Reports 10 1751 PMID: 29779899
Levardon et al (2018) Expansion of Airway Basal Cells and Generation of Polarized Epithelium. Bio Protoc 8 PMID: 30009215
McHugh et al (2018) Preclinical comparison of proteasome and ubiquitin E1 enzyme inhibitors in cutaneous squamous cell carcinoma: the identification of mechanisms of differential sensitivity. Oncotarget 9 20265 PMID: 29755650
Hepburn et al (2018) Targeting the spliceosome for cutaneous squamous cell carcinoma therapy: a role for c-MYC and wild-type p53 in determining the degree of tumour selectivity. Oncotarget 9 23029 PMID: 29796170
Ng-Blichfeldt et al (2018) Retinoic acid signaling balances adult distal lung epithelial progenitor cell growth and differentiation. EBioMedicine 36 461 PMID: 30236449
McKown et al (2018) Impaired adhesion of induced pluripotent stem cell-derived cardiac progenitor cells (iPSC-CPCs) to isolated extracellular matrix from failing hearts. Heliyon 4 e00870 PMID: 30364772
Pirie et al (2018) Mouse genome-wide association studies and systems genetics uncover the genetic architecture associated with hepatic pharmacokinetic and pharmacodynamic properties of a constrained ethyl antisense oligonucleotide targeting Malat1. PLoS Genet 14 e1007732 PMID: 30372444
Rath et al (2018) Rho Kinase Inhibition by AT13148 Blocks Pancreatic Ductal Adenocarcinoma Invasion and Tumor Growth. Cancer Res 78 3321 PMID: 29669760
Ford et al (2018) A Cell/Cilia Cycle Biosensor for Single-Cell Kinetics Reveals Persistence of Cilia after G1/S Transition Is a General Property in Cells and Mice. Dev Cell 47 509 PMID: 30458140
Sheng et al (2018) A stably self-renewing adult blood-derived induced neural stem cell exhibiting patternability and epigenetic rejuvenation. Nat Commun 9 4047 PMID: 30279449
Ackermann et al (2018) Bioreactor-based mass production of human iPSC-derived macrophages enables immunotherapies against bacterial airway infections. Nat Commun 9 5088 PMID: 30504915
Pitha et al (2018) Rho-Kinase Inhibition Reduces Myofibroblast Differentiation and Proliferation of Scleral Fibroblasts Induced by Transforming Growth Factor β and Experimental Glaucoma. Transl Vis Sci Technol 7 6 PMID: 30479877
Vu et al (2018) Neural stem cells for disease modeling and evaluation of therapeutics for Tay-Sachs disease. Orphanet J Rare Dis 13 152 PMID: 30220252
Tidball et al (2018) Generating Loss-of-function iPSC Lines with Combined CRISPR Indel Formation and Reprogramming from Human Fibroblasts. Bio Protoc 8 PMID: 30320153
Mou et al (2016) Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells Cell: Stem Cell 19 217 PMID: 27320041
Foulke-Abel et al (2016) Human Enteroids as a Model of Upper Small Intestinal Ion Transport Physiology and Pathophysiology. Nat Commun 150 638 PMID: 26677983
Guye et al (2016) Genetically engineering self-organization of human pluripotent stem cells into a liver bud-like tissue using Gata6. Sci Rep 7 10243 PMID: 26732624
Balcioglu et al (2016) Tumor-induced remote ECM network orientation steers angiogenesis. Development 6 22580 PMID: 26931404
Wilson et al (2016) Cryopreservation of Brain Endothelial Cells Derived from Human Induced Pluripotent Stem Cells Is Enhanced by Rho-Associated Coiled Coil-Containing Kinase Inhibition. Tissue Eng Part C Methods 22 1085 PMID: 27846787
Neely et al (2012) DMH1, a highly selective small molecule BMP inhibitor promotes neurogenesis of hiPSCs: comparison of PAX6 and SOX1 expression during neural induction. Proc Natl Acad Sci U S A 3 482 PMID: 22860217
Guo et al (2012) Slug and Sox9 cooperatively determine the mammary stem cell state. J Cell Sci 148 1015 PMID: 22385965
Walker et al (2012) Intra-axonal translation of RhoA promotes axon growth inhibition by CSPG. J Cell Sci 32 14442 PMID: 23055514
Lee et al (2011) The Relaxing Effect of α-Defensin 1 on the Adrenergic Responses of Rat Bladder. Dev Biol 15 143 PMID: 21860592
Vallenius et al (2011) An association between NUAK2 and MRIP reveals a novel mechanism for regulation of actin stress fibers. Integr Biol (Camb) 124 384 PMID: 21242312
Hartmann et al (2005) CXCR4 chemokine receptor and integrin signaling co-operate in mediating adhesion and chemoresistance in small cell lung cancer (SCLC) cells. Oncogene 24 4462 PMID: 15806155
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Schneider et al (2004) Signal transduction underlying CB-induced contraction of human urinary bladder. J Biol Chem 309 1148 PMID: 14769832
Guillemot and Citi (2006) Cingulin regulates claudin-2 expression and cell proliferation through the small GTPase RhoA. Mol Biol Cell 17 3569 PMID: 16723500
Wyckoff et al (2006) ROCK- and myosin-dependent matrix deformation enables protease-independent tumor-cell invasion in vivo. Curr Biol 16 1515 PMID: 16890527
Rajnicek et al (2006) Temporally and spatially coordinated roles for Rho, Rac, Cdc42 and their effectors in growth cone guidance by a physiological electric field. J Cell Sci 119 1723 PMID: 16595546
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Bhadriraju et al (2007) Quantifying myosin light chain phosphorylation in single adherent cells with automated fluorescence microscopy. J Biol Chem 8 43 PMID: 17941977
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Ruiz and Chen (2008) Emergence of patterned stem cell differentiation within multicellular structures. J Cell Sci 26 2921 PMID: 18703661
Inan and Büyükafşar (2008) Antiepileptic effects of two Rho-kinase inhibitors, Y-27632 and fasudil, in mice. Am J Pathol 155 44 PMID: 18536751
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Reviews for Y-27632 dihydrochloride
Average Rating: 5 (Based on 13 Reviews.)
Have you used Y-27632 dihydrochloride?
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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.