Rock inhibitor y 27632

Manufactured by MedChemExpress

Sourced in United States

ROCK Inhibitor y-27632 is a potent and selective Rho-associated protein kinase (ROCK) inhibitor. It inhibits ROCK1 and ROCK2 with IC50 values of 0.14 μM and 0.3 μM, respectively. ROCK Inhibitor y-27632 has been widely used as a research tool to study the roles of ROCK in various cellular processes.

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2 protocols using rock inhibitor y 27632

Adipose-Derived Stem Cell Differentiation

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ASCs were seeded at 10.2 × 10⁴ ASC per cm² in a 35-mm culture dish (Corning, Corning, NY). The ASCs were cultured with RoosterNourish™-MSC-XF culture media (RoosterBio, Inc., Ballenger Creek, Maryland). The differentiation of the cell sheets started when they reached confluence, on day 4 after initial seeding.

- Undifferentiated ASCCSs: ASCs were cultured with RoosterNourish™-MSC-XF. The culture media were replaced every 2 days, up to 18 days from the initial seeding day.

- Osteoblast cell sheets: ASCs were cultured with Osteomax-XF Differentiation Medium (Millipore-Sigma, Burlington, MA). The culture media were replaced every 3 days, up to 17 days from the initial seeding day.

- Chondrocyte cell sheets: ASCs were cultured with a MesenCult™-ACF Chondrogenic Differentiation Kit (Stem Cell, Vancouver, Canada). The culture media were replaced every 2 days, up to 19 days from the initial seeding, and were also replaced when 10 M of ROCK Inhibitor y-27632 (MedChemExpress, Monmouth Junction, NJ, United States) was used.

- Skeletal muscle differentiation medium (PromoCell, Heidelberg, Germany) was used to show that the cells do not form a multilayer cell sheet and the cell density does not change by measuring the transmittance.

Ochiai J., Villanueva L., Niihara H., Niihara Y, & Oliva J. (2022). Posology and Serum-/Xeno-Free Engineered Adipose Stromal Cells Cell Sheets. Frontiers in Cell and Developmental Biology, 10, 873603.

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Inhibition of Fibrosis Pathways

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HE, PAS and Masson's trichrome staining kits were purchased from Nanjing Jiancheng Biology Engineering Institute (Nanjing, Jiangsu, China). Dulbecco's modified Eagle's medium (DMEM), foetal bovine serum (FBS), penicillin and streptomycin were purchased from GIBCO BRL. Thrombin and dexamethasone powder were from Sigma‐Aldrich Co. Dabigatran was obtained from Boehringer Ingelheim Pharma. Inhibitors used were PAR1 antagonist SCH79797 (Tocris Bioscience), RhoA inhibitor Y‐16, ROCK inhibitor Y‐27632, YAP inhibitor verteporfin (MedChemExpress) and the actin polymerization inhibitor Cytochalasin D (Abcam). The antibodies used were anti‐collagen I, anti‐CTGF, anti‐β‐actin, anti‐GAPDH (Biosynthesis Biotechnology); anti‐RhoA, anti‐ROCK, anti‐phosphorylated MLC2, anti‐MLC2, anti‐phosphorylated SMAD2, anti‐SMAD2, anti‐phosphorylated YAP, anti‐α‐SMA, anti‐cyclin D1 (Cell Signaling Technology) and anti‐YAP (Santa Cruz Biotechnology).

Deng Z., Xie H., Cheng W., Zhang M., Liu J., Huo Y., Liao Y, & Cheng Y. (2020). Dabigatran ameliorates airway smooth muscle remodeling in asthma by modulating Yes‐associated protein. Journal of Cellular and Molecular Medicine, 24(14), 8179-8193.

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