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Y-27632 - a ROCK inhibitor with a valuable role in stem cell biology


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].

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].


Y-27632 dihydrochlorideY-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 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].

More products for stem cell research

Y-27632 dihydrochloride is available from Hello Bio at low prices - up to 50% less than other suppliers. It joins our range of low cost small molecule modulators of stem cell processes and biology. Browse our range of small molecule stem cell modulators below:


1, Amano M et al (2010) Rho-kinase/ROCK: A key regulator of the cytoskeleton and cell polarity. Cytoskeleton (Hoboken) 67(9) 545-54. Pubmed ID:20803696

2, Riento K et al (2003) Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 4(6) 446-56. Pubmed ID:12778124

3, Wu Y et al (2016) ROCK inhibitor Y27632 promotes proliferation and diminishes apoptosis of marmoset induced pluripotent stem cells by suppressing expression and activity of caspase-3. Theriogenology 85(2) 302-14. Pubmed ID:26476594

4, Kourdougli N et al (2015) Detrimental effect of post Status Epilepticus treatment with ROCK inhibitor Y-27632 in a pilocarpine model of temporal lobe epilepsy. Front Cell Neurosci 9 413 Pubmed ID:26557054

5, Breitenlechner C et al (2003) Protein kinase A in complex with Rho-kinase inhibitors Y-27632, Fasudil, and H-1152P: structural basis of selectivity. Structure 11(12) 1595-607. Pubmed ID:14656443

6, Jung DW et al (2014) Reprogram or reboot: small molecule approaches for the production of induced pluripotent stem cells and direct cell reprogramming. ACS Chem Biol 9(1) 80-95. Pubmed ID:24245936

7, Watanabe K et al (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25(6) 681-6. Pubmed ID:17529971

8, Ohgushi M et al (2010) Molecular pathway and cell state responsible for dissociation-induced apoptosis in human pluripotent stem cells. Cell Stem Cell 7(2) 225-39. Pubmed ID:20682448

9, Lamas NJ et al (2015) Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells. Stem Cells Cloning 8 15-26. Pubmed ID:25609984

10, Lee et al (2015) Effect of Rho-kinase Inhibitor, Y27632, on Porcine Corneal Endothelial Cell Culture, Inflammation and Immune Regulation. Ocul Immunol Inflamm 16 42005 Pubmed ID:26471144

11, Kurosawa H (2012) Application of Rho-associated protein kinase (ROCK) inhibitor to human pluripotent stem cells. J Biosci Bioeng 114(6) 577-81. Pubmed ID:22898436

12, Martin-Ibañez R et al (2008) Novel cryopreservation method for dissociated human embryonic stem cells in the presence of a ROCK inhibitor. Hum Reprod 23(12) 2744-54. Pubmed ID:18716037