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 (Cat. No. 04-0012-02).

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

Specifications

Product Name: Stemolecule Y27632

Catalog Number: 04-0012

Size:

• 2 mg (Cat. No. 04-0012)
• 10 mg (Cat. No. 04-0012-10)
• 2 mg in 10 mM DMSO (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.

Documents

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

References

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).
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• 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).
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• 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).
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• 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).
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• 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).
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• 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).
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• 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).
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