Y-27632 (dihydrochloride) -ROCK Inhibitor

Y-27632 is a cell-permeable small molecule that selectively and potently inhibits Rho-associated coiled-coil containing protein kinases (ROCK1 and ROCK2). By competitively binding the ATP catalytic site of ROCK enzymes, Y-27632 effectively suppresses their kinase activity (ROCK1 Ki = 220 nM; ROCK2 Ki = 300 nM) (Ishizaki et al.,2000; Davies et al.,2000). This selective inhibition modulates cytoskeletal dynamics, cell adhesion, and apoptosis pathways, making Y-27632 a valuable tool in cellular biology and regenerative medicine research.

Key Applications of Y-27632

Maintenance and Self-Renewal of Stem Cells

Enhances Cell Survival:

Y-27632 significantly improves survival rates of human embryonic stem (ES) cells after single-cell dissociation by preventing apoptosis triggered by cell detachment (anoikis). This effect increases cloning efficiency and facilitates expansion of ES cultures (Watanabe et al., 2007).

Supports Embryoid Body Formation

When used in forced-aggregation protocols, Y-27632 enhances the formation of embryoid bodies, promoting three-dimensional differentiation potential (Ungrin et al., 2008).

Improves Post-Thaw Viability

The compound protects cryopreserved single human ES cells from apoptosis after thawing, resulting in higher recovery and viability (Martin-Ibañez et al., 2008, Li et al., 2008, Li et al., 2009).

Neural Precursor Survival

In embryonic stem cell-derived neural precursor cells, Y-27632 blocks apoptosis following cell dissociation and transplantation, supporting successful cell engraftment (Koyanagi et al., 2008).

Promotes Neurite Outgrowth

Y-27632 regulates microtubule remodeling to regulate neurite outgrowth (Chen et al., 2013).

Reprogramming and Lineage Conversion

Y-27632 serves as a critical component in direct lineage reprogramming protocols. In combination with other small molecules (such as CHIR99021, RepSox, Forskolin, SP600125, Gö6983, and Valproic Acid), it facilitates the conversion of fibroblasts into mature neurons, broadening its utility in regenerative medicine and cellular engineering (Hu et al., 2015).

Differentiation Protocols

During initiation of differentiation, particularly in human ES cell monolayers, Y-27632 enhances cell survival, enabling more efficient and reproducible differentiation outcomes (Rezania et al., 2014). Additionally, by modulating actomyosin contractility in the stem cell niche, Y-27632 influences cell shape and mechanical cues that regulate stem cell lineage commitment (McBeath et al., 2004).

Y-27632 also improves viability and proliferation in other cell types, such as human keratinocytes cultured in vitro, enabling their efficient immortalization and long-term culture (Chapman et al., 2010).

Y-27632 (GMP-compliant)

This product is complied with ICH-Q7 (GMP for APIs) and is intended as a raw material for the commercial production of regenerative medicine products. It has been tested for viable count, as well as bacterial endotoxins, mycoplasma contamination, and residual solvent.

• GMP compliant
• Process validation analytical validation carried out
• No animal-derived ingredients are used as raw materials

Y-27632, MF

This is an ISO9001-compliant product developed for the commercial manufacturing of regenerative medicine products. It has been tested for viable count, as well as bacterial endotoxins and mycoplasma contamination.
The management and manufacturing standards have been established internally.

• ISO9001 compliant
• Reduction of contamination risk through the use of dedicated equipment for manufacturing culture media additives
• Process validation, analytical validation, and cleaning validation carried out
• No animal-derived ingredients are used as raw materials

CultureSure^® Y-27632

This product was developed for basic research in drug discovery and regenerative medicine. It has been tested for bacterial endotoxins and mycoplasma contamination.

• ISO9001 compliant
• No animal-derived ingredients are used as raw materials

CultureSure^® 10 mmol/L Y-27632 Solution, Animal-derived-free

This product is a 10 mmol/L aqueous solution of Y-27632. It has been tested for sterility, as well as bacterial endotoxins and mycoplasma contamination. It has been 0.1 μm filter-sterilized and can be added directly to the culture medium.

• ISO9001 compliant
• No animal derived-ingredients are used as raw materials

Here is a protocol for passaging, freezing, and thawing human ES/iPS cells (single cells), using our product Y-27632.

Y-27632 is used for effective differentiation induction of iPS/ES cells.
Based on the latest findings reported in the literature, we introduce examples of the usage steps for our products.

Nerves

References

• Vihma, H. et al.: Nat Commun., 15, 5558 (2024).
• Yusuke, H. et al.: Front Cell Dev Biol., 13, 1536947 (2025).

Retina

References

• Kanda. GN. et al.: Elife, 11, e77007 (2022).

Lung

References

• Yuki, K. et al.: J Gen Virol. 106, 002131 (2025).
• Rina H. et al.: Life Sci Alliance. 8, e202402837 (2025).
• Masatoshi, K. et al.: J Virol. 99, e01853-24 (2024).
• Hiroki, F.et al.: iScience. 27, 109597(2024).

Kidney

References

• Tsujimoto, H. et al.: Cell Reports, 31, 107476 (2020).

* Click to move to the PubMed Central^® search results.* Click to move to the PubMed Central^® search results.

The powder Y-27632 product has been confirmed to be soluble in water. After dilution, filter sterilize before use. Refer to the protocol at the top of this page for general usage in ES/iPS cell culture.

Amount of solvent when preparing solution

Literature and Education

• Map of Culture Compounds and Cytokines
• Small Molecules Catalog
• On-demand webinar：Impact of ROCK inhibitor in suspension aggregate culture of iPS cells
• FUJIFILM Wako Life Science Webinar Series

Publications