ROCK inhibitor Y-27632

(HB2297)
Technical documents: SDS CoA Datasheet

Product overview

Name ROCK inhibitor Y-27632
Description Y-27632 is a selective ROCK inhibitor. Frequently used 3D growth matrix component and for production of organoids. Also used when inducing neurons from fibroblasts.
Biological description

The ROCK inhibitor Y-27632 is a selective and cell permeable inhibitor of the Rho-associated protein kinase (ROCK) (Ki values are 0.14, 26 ,25 and >250 µM at p160ROCK, PKC, PKA and MLCK respectively).


Y-27632 has many biological actions and is an important small molecule modulator of stem cells. It diminishes hESC and hiPSC dissociation-induced apoptosis, enhances survival and colony formation of dissociated hESC without affecting pluripotency or self-renewal.


Y-27632 also improves hESC and iPSCs postthaw viability and survival rate during cryopreservation.


Additionally, Y-27632 inhibits smooth muscle contractility and shows antihypertensive effect. 

It is frequently used as a 3D growth matrix component and for production of organoids (e.g. brain organoids).

Alternative names Y27632 dihydrochloride
Purity >99%
Customer comments

The ROCK inhibitor Y-27632 is working as expected. Verified customer, University of Louisville

Worked well just as it should do. Kept me informed of when product would be delivered and notified me when delivered. Good packaging and data sheet. Easy to dissolve and use and it worked! Verified customer, University of Cardiff

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Biological Data

Application notes

Introduction

Rho kinase (Rho-associated coiled coil protein kinase or ROCK) is a major downstream effector for the small GTP-binding protein Rho [1]. Rho kinase is involved in various biological processes including cell mitosis adhesion, cytoskeletal adjustments, muscle cell contraction, tumor cell invasion and a series of cell biological phenomena [2]. Through a series of cascading effects, Rho kinase influences actin and microtubule regulation of apoptosis and thus plays a key role in early embryonic development [3].The Rho kinase signalling pathway is highly activated in many pathological conditions [4].

Rock inhibitor Y-27632 is a Rho kinase inhibitor which selectively inhibits p160ROCK (ROCK1) with a Ki value of 140 nM and binds to the Rho kinase ATP binding pocket in an ATP-competitive manner [5].


Uses

Y-27632 has been shown to have a diverse range of actions and has been used in a variety of biological systems. It is the most commonly used of the ROCK inhibitors in stem cell research and is used as a multifunctional reagent in many stem cell related processes [6]. Y-27632 demonstrates many positive effects across various cell types, in particular human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) [7] and has greatly improved a number of practical stem cell procedures [8]. For example it has been shown that Y-27632 increases cell adhesion and proliferation and increases post-thaw cell survival and post-passaging cell viability [9,10].


Cellular dissociation

Cell dissociation is one of the most common manipulations in stem cell research. During cellular dissociation both hESC and hIPSCs are vulnerable to dissociation-induced apoptosis or anoikis (a form of apoptosis induced by inappropriate cell-cell or cell-ECM interactions) [3,11]. Y-27632 is of particular interest as it allows hPSCs to escape this dissociation-induced apoptosis. Y-27632 prevents apoptosis of dissociated hPSCs and increases their survival rate and plating efficiency [11].


Cryopreservation

Cryopreservation is also an important part of stem cell research. Y-27632 improves the recovery of cryopreserved hPSCs and has greatly helped simplify the cryopreservation procedure. The slow-freezing and rapid-thawing procedure which uses DMSO as a cryoprotectant is commonly used in stem cell cryopreservation, however this method is not suitable for hPSCs, therefore vitrification (fast freezing, slow thawing) is used [12]. Vitrified hPSCs do however still suffer from high levels of cell death and do not passage well as single cells [11]. The addition of Y-27632 to freezing and post-thawing medium enhances colony formation efficiency and treatment with Y-27632 during cryopreservation also increases survival rate and cell adhesion of freeze-thawed dissociated hES [12].

Solubility & Handling

Storage instructions Room temperature (desiccate)
Solubility overview Soluble in water (100mM), PBS (100mM), DMSO (100mM)
Handling
  • If using the compound as a cell culture supplement, you should dilute the stock solution into culture medium immediately before use.
  • Cell culture media can be prewarmed before adding the compound to avoid potential precipitation of the compound.
  • Y-2762 is often removed from medium within 20 hours
  • The compound has been shown to be effective at a final concentration of 10 µM (Ungrin et al.; Watanabe et al.)
  • Mix and filter supplemented media using a 0.2 µm low‑protein binding filter.
  • Please avoid a final DMSO concentration above 0.1% due to potential cell toxicity
Important This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use.

Calculators

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Dilution

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Chemical Data

Chemical name trans-4-[(1R)-1-Aminoethyl]-N-4-pyridinylcyclohexanecarboxamide dihydrochloride
Molecular Weight 320.26
Chemical structure Product image
Molecular Formula C14H21N3O.2HCl
CAS Number 129830-38-2
PubChem identifier 9901617
SMILES C[C@H](C1CCC(CC1)C(=O)NC2=CC=NC=C2)N.Cl.Cl
Source Synthetic
InChi InChI=1S/C14H21N3O.2ClH/c1-10(15)11-2-4-12(5-3-11)14(18)17-13-6-8-16-9-7-13;;/h6-12H,2-5,15H2,1H3,(H,16,17,18);2*1H/t10-,11?,12?;;/m1../s1
InChiKey IDDDVXIUIXWAGJ-DDSAHXNVSA-N
MDL number MFCD03490488
Appearance White solid

References for ROCK inhibitor Y-27632

References are publications that support the biological activity of the product
  • 1, Rho-kinase/ROCK: A key regulator of the cytoskeleton and cell polarity.

    Amano et al (2010) Cytoskeleton (Hoboken) 67(9) : 545-54
  • 2, Rocks: multifunctional kinases in cell behaviour.

    Riento and Ridley (2003) Nat Rev Mol Cell Biol 4(6) : 446-56
  • 3, ROCK inhibitor Y27632 promotes proliferation and diminishes apoptosis of marmoset induced pluripotent stem cells by suppressing expression and activity of caspase 3.

    Wu et al (2016) Theriogenology 85(2) : 302-14
  • 4, Detrimental effect of post Status Epilepticus treatment with ROCK inhibitor Y-27632 in a ... model of temporal lobe epilepsy.

    Kourdougli et al (2015) Front Cell Neurosci. 9 : 413
  • 5, Protein kinase A in complex with Rho-kinase inhibitors Y-27632, Fasudil, and H-1152P: structural basis of selectivity.

    Breitenlechner et al (2003) Structure 11(12) : 1595-607
Publications
These publications cite the use of ROCK inhibitor Y-27632 purchased from Hello Bio:
  • Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell and Testis Development Research.

    Pryzhkova, M.V., Jordan, P.W. (2019) Tissue Eng Regen Med : 29(02)
    PubMedID: 32114677
  • An automatic cell culture platform for differentiation of human induced pluripotent stem cells

    Pitingolo et al (2020) Microelectronic Engineering : 111371
  • The generation of an induced pluripotent stem cell line (DCGi001-A) from an individual with FOXG1 syndrome carrying the c.460dupG (p.Glu154fs) variation in the FOXG1 gene

    Clarket et al (2020) Stem Cell Res. 49 : 102018
    PubMedID: 33096386
  • Adaptation of the AID system for stem cell and transgenic mouse research

    Jordan et al (2020) Stem Cell Res. : 102078
  • Neuroepithelial organoid patterning is mediated by Wnt-driven Turing mechanism

    Fattah et al (2021) bioRxiv Preprint : https://www.biorxiv.org/content/

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