Stemolecule™ Y27632

Y27632 is a cell-permeable small molecule Rho-associated kinase (ROCK) inhibitor¹. Y27632 has been found to prevent apoptosis as well as enhance the survival and cloning efficiency of dissociated human embryonic stem (ES) cells without affecting their self-renewal properties or pluripotency². This molecule has also been shown to enhance survival during the transplantation of ES cell-derived neural precursors³. Y27632 in combination with Pifithrin-u significantly improves cell recovery after cryopreservation⁴.

Stemolecule Y27632 is also available as a 10 mM DMSO Stock Solution in DMSO (2 mg in 624.4 μl) (Cat. No. 04-0012-02).

Y27632 is widely used in naive stem cell protocols⁵, such as the 5i/L/A/ protocol.

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Stemgent and the Stemolecule brand name are trademarks of REPROCELL Inc., Japan.

Product Name: Stemolecule Y27632

Catalog Number: 04-0012

Size:

• 2 mg (Cat. No. 04-0012)
• 10 mg (Cat. No. 04-0012-10)
• 2 mg solution in 10 mM DMSO (2 mg in 624.4 μl) (Cat. No. 04-0012-02)

Alternate Name(s):

• Y-27632
• (1R,4R)-4-((R)-1-aminoethyl)-N-(pyridine-4-yl)cyclohexanecarboxamide dihydrochloride

Chemical Formula: C₁₄H₂₁N₃O·2HCl

Molecular Weight: 320.3

CAS Number: 146986-50-7

Purity: Greater than 97 % by HPLC analysis

Formulation: White solid

Solubility: Reconstitute in DMSO to the desired concentration. For reconstitution instructions please reference product specifications sheet.

Storage and Stability: Store powder at 4 °C protected from light. Following reconstitution, store aliquots at −20 °C. Stock solutions are stable for 6 months when stored as directed.

Quality Control: The purity of Y27632 was determined by HPLC analysis. The accurate mass was determined by mass spectrometry. Cellular toxicity of Y27632 was tested on mouse embryonic stem cells.

Specification Sheets:

• 04-0012 Specifications Sheet
• 04-0012-10 Specifications Sheet
• 04-0012-02 Specifications Sheet

Safety Data Sheets:

• 04-0012 and 04-0012-10 Safety Data Sheet
• 04-0012-02 Safety Data Sheet

1. Ishizaki, T., Uehata, M., Tamechika, I., Keel, J., Nonomura, K., Maekawa, M., and Narumiya, S. Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol Pharmacol 57: 976-983 (2000).
2. Watanabe, K., Ueno, M., Kamiya, D., Nishiyama, A., Matsumura, M., Wataya, T., Takahashi, J.B., Nishikawa, S., Nishikawa, S., Muguruma, K., and Sasai, Y. A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25: 681-686 (2007).
3. Koyanagi, M., Takahashi, J., Arakawa, Y., Doi, D., Fukuda, H., Hayashi, H., Narumiya, S., and Hashimoto, N. Inhibition of the Rho/ROCK pathway reduces apoptosis during transplantation of embryonic stem cell-derived neural precursors. J Neurosci Res 86: 270-280 (2008).
4. Xu, X., Cowley, S., Flaim, C.J., James, W., Seymour, L.W., Cui, Z. Enhancement of cell recovery for dissociated human embryonic stem cells after cryopreservation. Biotechnol Prog 26: 781-788 (2010).
5. Fonseca, SAS; Costas, RM; Pereira, LV. Searching for naive human pluripotent stem cells. World Journal of Stem Cells, 7(3), 649 (2015).

Additional Publications

• Li Y; Li J; Wang J; Zhang S; Giles K; Prakash TP; Rigo F; Napierala JS; Napierala M. Premature transcription termination at the expanded GAA repeats and aberrant alternative polyadenylation contributes to the Frataxin transcriptional deficit in Friedreich’s ataxia. Human Molec Genetics 31:3539 (2022).
• Cohen B; McPhie D; Ren B; Garcia-Leon J; Verfaille C. SOX10 Expression Levels may be a Critical Mediator of White Matter Deficits in Schizophrenia. BioRivX https://doi.org/10.21203/rs.3.rs-1808873/v1: (2022).
• Yang D; Patel S; Szlachcic J; Scaduto S; Putluri N; Sreekumar A; Suliburk J; Metzker M; Balasubramanyam A; Borowiak M. Pancreatic Differentiation of Stem Cells Reveals Pathogenesis of a Syndrome of Ketosis-Prone Diabetes. Diabetes 70:2419 (2021).
• Facoioli R; Lojudice FH; Anauate AC; Maquigussa E; Nishiura JL; Heilberg IP; Sogayar MC; Boir MA. Kidney organoids generated from erythroid progenitors cells of patients with autosomal dominant polycystic kidney disease. PLOS One 16:e0252156 (2021).
• Wang T; Liu H; Itoh K; Oh S; Zhao L ;Murata D; Sesaki H; Hartung T; Na CH; Wang J. C9orf72 regulates energy homeostasis by stabilizing mitochondrial complex I assembly. Cell Metabolism 33:531-546.e9 (2021).
• Jiang Y; Sun Si0J; Wei R; Zhang N; Liao S-Y; Tse H-F. Myocardial repair of bioengineered cardiac patches with decellularized placental scaffold and human-induced pluripotent stem cells in a rat model of myocardial infarction. Stem Cell Res Therap 12:13 (2021).
• Carberry BJ; Hergert JE; Yavitt FM; Hernandez JJ; Speckl KF; Bowman CN; McLeod RR; Anseth KS. 3D printing of sacrificial thioester elastomers using digital light processing for templating 3D organoid structures in soft biomatrices. Biofabrication 13:044104 (2021).
• Wen W; Quan Z-J; Li S-A; Yang Z-X; Fu Y-W; Zhang F; Li G-H; Zhao M; Yin M-D; Xu J; Zhang J-P; Cheng T; Zhang X-B. Effective control of large deletions after double-strand breaks by homology-directed repair and dsODN insertion. Genome Biology 22:236 (2021).
• Schoger E; Zimmermanm W-H; Cyganek L Zalarayán LC. Establishment of a second generation homozygous CRISPRa human induced pluripotent stem cell (hiPSC) line for enhanced levels of endogenous gene activation. Stem Cell Res 56:102518 (2021).
• Huang X; Park K; Gontarz P Zhang B; Pan J; McKenzie Z; Fischer LA; Dong C; Dietmann S; Xing X; Shliaha PB; Yang J; Li D ;Ding J; Lungjangwa T; Mitalipova M; Khan S; Imsoonthornruksa S; Jensen M; Wang T; Kadoch C; Raenisch R; Wang J; Theunissen TW. OCT4 cooperates with distinct ATP-dependent chromatin remodelers in naïve and primed pluripotent states in human. Nature Commun 12:5123 (2021).
• Bakare AB; Meshrkey F; Lowe B; Molder C; Rao RR; Zhan J; Iyer S. MitoCellPhe reveals mitochondrial morphologies in single fibroblasts and clustered stem cells. Am J Phys Cell Physiol 321:C735-748 (2021).
• Rajput SA; Liang S-J; Wang W-Q; Yan H-C. Lycopene Protects Intestinal Epithelium from Deoxynivalenol-Induced Oxidative Damage via Regulating Keap1/Nrf2 Signaling. Antioxidants 10:1493 (2021).
• Meshrkey F; Ayuso AC; Rao RR; Iyer S. Quantitative analysis of mitochondrial morphologies in human induced pluripotent stem cells for Leigh syndrome. Stem Cell Res 57:102572 (2021).
• Bhan A; Ansari K; Chen MY; Jandial R. Human induced pluripotent stem cell-derived platelets loaded with lapatinib effectively target HER2+ breast cancer metastasis to the brain. Sci Rep 11:16866 (2021).
• Vershkovv D; Yilmaz A; Yanuka O; Nielsen A: Benvenisty N. Genome-wide screening for genes involved in the epigenetic basis of fragile X syndrome. Stem Cell Rep 17:1048 (2022).
• Dong C; Fu S; Karvas RM; Chew B; Fischer LA; Xing X; harrison JK; popli P; Kommagani R; Wang T; Zhang B; Theunissen TW. A genome-wide CRISPR-Cas9 knockout screen identifies essential and growth-restricting genes in human trophoblast stem cells. Nature Commun 13:2548 (2022).
• Michielin F; Giobbe GG; Luni C; Hu Q; Maroni I; Ofrord MR; Manfredi A; Di Filipp9; David AL; Cacchiarelli D; De Coppi P Eaton S; Elvassore N. The Microfluidic Environment Reveals a Hidden Role of Self-Organizing Extracellular Matrix in Hepatic Commitment and Organoid Formation of hiPSCs. Cell Rep 33:108453 (2020).
• Ryan SK; Jordan-Sciutto KL; Anderson SA. Protocol for Tri-culture of hiPSC-Derived Neurons, Astrocytes, and Microglia. STAR Protocols 1:100190 (2020).
• Akiyama H; Jalloh S; Park S; Lei M; Mostoslavski G; Gummuluru S. Expression of HIV-1 intron-containing RNA in Microglia Induces Inflammatory Responses. J Virol in press:doi:10.1128/JVI.01386-20 (2020).
• Barbar L; Rusielewica T; Zimmer M; Kalpana K; Fossati V. Isolation of Human CD49f+ Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays. STAR Protocols :doi.org/10.1016/j.xpro.2020.100172 (2020).
• Scoger E; Argyiou L; Cyganke L; Zelarayán. Generation of homozygous CRISPRa human induced pluripotent stem cell (hiPSC) lines for sustained endogenous gene activation. Stem Cell Res :101944 (2020).
• Wong C-K; Luk HK-H; Lai W-H; Lau Y-M; Zhang RR; Wong AC-P; Lo GC-S; Chan K-H; Hung IF-N; Tse H-F; Woo PC-Y; Lau SK-P; Siu C-W. Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Platform to Study SARS-CoV-2 Related Myocardial Injury. Circulation J :doi:10.1253/circj.CJ-20-0881 (2020).
• Ambrosini YM; Park Y; Jergens AE; Shin W; Min S; Atherly T; Borcherding DC; Jang J; Allenspach K; Mochel JP; Kim HJ. Recapitulation of the accessible interface of biopsy-derived canine intestinal organoids to study epithelial-luminal interactions. PLoS ONE 15:e231423 (2020).
• Chu J; Fitzgerald ML; Sehgal M; Manley W; Fitzgerald S; Naung H; Goldberg EM; Anderson SA. Enhanced maturation of human stem cell derived interneurons by mTOR activation . bioRxiv :dx.doi.org/10.1101/777714 (2019).
• Chen D; Sun N; Hou L; Kim R; Faith J; Aslanyan M; Tao Y; Zheng Y; Fu J; Liu W; Kellis M; Clark A. Human Primordial Germ Cells Are Specified from Lineage-Primed Progenitors. Cell Rep 29:4568-4582.e5 (2019).
• Koutmani Y; Gampierakis IA; Polissidis A; Ximerakis M: Koutsoudaki PN; Polyzos A; Agrogiannis G; Karaliota S; Thomaidou D; Rubin LL; Politis PK; Karakis KP. CRH Promotes the Neurogenic Activity of Neural Stem Cells in the Adult Hippocampus. Stem Cell Rep 28:932-945.e7 (2019).
• Nagaraja S; Quezada MA; Gillespie SM; Arzt M; Lennon JJ; Woo PJ; Hovestadt V; Kambhampati M; Filbin MG; Suva ML; Nazarian J; Monje M. Histone Variant and Cell Context Determine H3K27M Reprogramming of the Enhancer Landscape and Oncogenic State. Molecular Cell 76:dev.138982 (2019).
• Trautsch I; Heta E; Soong PL; Levent E; Nikolaev VO; Bogeski I ; Katschinski DM; Mayr M; Zimmermann W-H. Optogenetic Monitoring of the Glutathione Redox State in Engineered Human Myocardium. Front Physiol 10:272 (2019).
• LI S; Zhang H; Wang A; Liu Y; Yue F; Abulati X; Zhang C; Li L. Differentiation of adult human retinal pigment epithelial cells into dopaminergic-like cells in vitro and in the recipient monkey brain. Molecular Med 25:9 (2019).
• Talbot NC; Shannon AE; Garrett WM. Pancreatic duct-like cell line derived from pig embryonic stem cells: Expression of uroplakin genes in pig pancreatic tissue. In vitro Cell Develop Biol - Animal 2019:1 (2019).
• Wadhwani AR; Affaneh A; Van Gulden S; Kessler JA. Neuronal Apolipoprotein E4 increases cell death and p‐tau release in Alzheimer's disease. Ann Neurol doi: 10.1002/ana.25455.: (2019).
• Kim JJ; Savas JN; Miller MT; Hu X; Carromeu C; Lavallée-Adam M; Freitas BCG; Muotri AR; Yates III JR; Ghosh A. Proteomic analyses reveal misregulation of LIN28 expression and delayed timing of glial differentiation in human iPS cells with MECP2 loss-of-function. Plos One 14:e0212553 (2019).
• Dai S-B; Shen T; Zheng T-T; Pu J-L; Chen X-Z. Plasmid-based generation of neural cells from human fibroblasts using non-integrating episomal vectors. Neural Regen Res 14:510-505 (2019).
• Scavuzzo MA; Chmielowiec J; Yang D; Wamble K; Chaboub LS; Duraine L; Tepe B; Glasgow SM; Arenkiel BR; Brou C; Deneen B; Borowiak M; . Pancreatic Cell Fate Determination Relies on Notch Ligand Trafficking by NFIA. Cell Reports 25:3811-3827.e7 (2018).
• Chen D; Liu W: Zimmerman J; Pastor WA; Kim R;  Hosohama L; Ho J; Aslanyan M; Gell JJ; Jacobsen SE; Clark AT. The TFAP2C-Regulated OCT4 Naive Enhancer Is Involved in Human Germline Formation. Cell Reports 25:3591-3602.e5 (2018).
• Worley KE; Chin AS; Wan LQ. Lineage-Specific Chiral Biases of Human Embryonic Stem Cells during Differentiation. Stem Cells Intl 2018:1848605 (2018).
• Kilander MB; Wang C-H; Chang C-H; Nestor JE; Herold K; Tsai J-W; Nestor MW; Lin Y-C. A rare human CEP290 variant disrupts the molecular integrity of the primary cilium and impairs Sonic Hedgehog machinery. Sci Rep 8:17335 (2018).
• Javed AO; Li Y; Muffat J; Su K-C; Cohen MA; Lungjangwa T; Auborg P; Cheeseman IM; Jaenisch R. Microcephaly Modeling of Kinetochore Mutation Reveals a Brain-Specific Phenotype. Cell Rep 25:368-382.e5 (2018).
• Memon B; Karam M; Al-Khawaga S; Abdelalim EM. Enhanced differentiation of human pluripotent stem cells into pancreatic progenitors co-expressing PDX1 and NKX6.1. Stem Cell Res Therap 9:15 (2018).
• Yucer N; Holzapfel M; Jenkins Vogel T; Lenaeus L; Ornelas L; Laury A; Sareen D; Barrett R; Karlan BY; Svendsen CN. Directed Differentiation of Human Induced Pluripotent Stem Cells into Fallopian Tube Epithelium. Sci Rep 7:10741 (2017).
• Lin VJT; Zolekar A; Shi Y; Koneru B; Dimitrijevich S; Di Pasqua AJ; Wang C-Y. Potassium as a pluripotency-associated element identified through inorganic element profiling in human pluripotent stem cells. Sci Reports 7:5005 (2017).
• Donegan JJ; Tyson JA; Branch SY; Beckstead MJ; Anderson SA; Lodge DJ. Stem cell-derived interneuron transplants as a treatment for schizophrenia: preclinical validation in a rodent model. Mol Psychiatry 22:1942-1501 (2017).
• McCracken KW; Aihara E; Martin B; Crawford CM; Broda T; Treguier J; Zhang X; Shannon JM; Montrose MH; Wells JM. Wnt / β-catenin promotes gastric fundus specification in mice and humans. Nature 541:182-187 (2017).
• Noack C; Haupt LP; Zimmerman W-H; Streckfuss-Bomeke; Zelarayan LC. Generation of a KLF15 homozygous knockout human embryonic stem cell line using paired CRISPR/Cas9n, and human cardiomyocytes derivation. Stem Cell Research 23:127-131 (2017).
• Badenes SM; Fernandes TG; Cordeiro CSM; Boucher S; Kuninger D; Vemuri MC; Diogo MM; Cabral JMS. Defined Essential 8 medium and vitronectin efficiently support xeno-free expansion of human induced pluripotent stem cells in stirred microcarrier culture systems. PLoS ONE 11:e0155296 (2016).
• Koehler KR; Ine J; Longworth-Mills E; Liu X-P; Lee J; Holt JR; Hashino E. Generation of inner ear organoids with functional hair cells from pluripotent stem cells. Nat Biotechnol 35:583-589 (2017).
• Du F; Chen C-H; Li Y; Hu Y; An L-Y; Yang L; Zhang J; Chen EY; Xu Jie. Derivation of rabbit embryonic stem cells from vitrified-thawed embryos. Cellular Reprog 17:453 (2015).
• DeRosa BA; Belle KC; Thomas BJ; Cukier HN; Pericak-Vance MA; Vance JM; Dykxhoom DM. hVGAT-mCherry: A novel molecular tool for analysis of GABAergic neurons derived from human pluripotent stem cells. Mol Cell Neurosci 68:244 (2015).
• Zimmerlan L; Park TS; Huo JS; Verma K; Pather SR; Talbot CC; Agarwal J; Steppan D; Zhang YW; Considine M; Guo H; Zhong X; Gutierrez C; Cope L; Canto-Soler MV; Friedman AD; Baylin SB; Zambdis ET. Tankyrase inhibition promotes a stable human naive pluripotent state with improved functionality. Development 2016:dev.138982 (2016).
• Tan H-K; Toh C-XDl Ma D; Yang B; Liu TM; Lu J; Wong C-W; Tan T-K; Li H; SYn C; Tan E-L; Lim B; Lim Y-P; Cook SA; Loh Y-H. Human finger-prick induced pluripotent stem cells facilitate the development of stem cell banking. Stem Cells Trans Medicine 3:586 (2014).
• Prè D; Nestor MW; Sproul AA; Jacob S; Koppensteiner P; Chinchalongpom V; Zimmer M; Yamamoto A; Noggle SA; Arancio O. A time course analysis of the electrophysiological properties of neurons differentiated from human induced pluripotent stem cells (iPSCs). PLoS ONE 9(7): e103418 (2014).
• Borenstrom C; Simonsson S; Enochson L; BIgdeli N; Branstsing C; Ellerstrom C; Hyllner J; Lindahl A. Footprint-Free Human Induced Pluripotent Stem Cells From Articular Cartilage With Redifferentiation Capacity: A First Step Toward a Clinical-Grade Cell Source. Stem Cells Translat Med 3:433-447 (2014).
• Youn H; Kim S-H; Choi KA; Kim S. Characterization of Oct4-GFP spermatogonial stem cell line and its application in the reprogramming studies. J Cell Biochem 114:920 (2013).
• Lian, X., Hsiao, C., Wilson, G, Zhu, K., Hazeltine, L.B., Azarin, S.M., Raval, K.K., Zhang, J., Kamp, T.J., and Palecek, S.P. (2012) Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci USA 109: E1848 (2012).