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 . 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 . 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 .The Rho kinase signalling pathway is highly activated in many pathological conditions .
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 .
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 . Y-27632 demonstrates many positive effects across various cell types, in particular human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs)  and has greatly improved a number of practical stem cell procedures . 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].
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 .
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 . Vitrified hPSCs do however still suffer from high levels of cell death and do not passage well as single cells . 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 .
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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:
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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
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