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Y 27632

Y 27632 is a Rho-associated protein kinase (ROCK) inhibitor that has been widely studied for its potential therapeutic applications.
This compound has demonstrated the ability to modulate various cellular processes, including cell migration, proliferation, and differentiation.
Researchers utilize Y 27632 to investigate its effects on a range of biological systems and disease models, such as neurological disorders, cardiovascular diseases, and cancer.
The detailed description and comparison of Y 27632 research protocols can help optimize experiments and lead to more informed decisions, streamlining the discovery process.
Pubcommpare.ai's AI-driven tools can assist researchers in locating relevant protocols from literature, preprints, and patents, and identify the best options using advanced comparisons.

Most cited protocols related to «Y 27632»

1
Reprogramming Fibroblasts to iPSCs
A schematic diagram of the procedure is described in Figure 4. Episomal plasmids and methods were described previously15 (link). Plasmid combination #19 (pEP4-E-O2S-E-T2K, pEP4-E-O2S-E-N2K and pCEP4M2L) was used for most reprogramming unless mentioned otherwise. Plasmids and EBNA mRNA were electroporated into fibroblast cells on Amaxa apparatus according to company instructions. One million cells were used in each electroporation, which were then plated into two 6-well plates. E8 + hydrocortisone media were used for the first 5–10 days, according to cell survival and proliferation after electroporation. When confluency was reached ~20%, hydrocortisone was removed. ES-like iPS cell colonies usually appear after ~25 days. Cells were then picked into individual wells with E8 (TGFβ or NODAL). Cells were passaged for ~ 15 passages before subcloning with Y27632 on Matrigel or vitronectin.
Chen G., Gulbranson D.R., Hou Z., Bolin J.M., Ruotti V., Probasco M.D., Smuga-Otto K., Howden S.E., Diol N.R., Propson N.E., Wagner R., Lee G.O., Antosiewicz-Bourget J., Teng J.M, & Thomson J.A. (2011). Chemically defined conditions for human iPS cell derivation and culture. Nature methods, 8(5), 424-429.
Publication 2011
Cells Cell Survival Electroporation Episomes Fibroblasts Hydrocortisone Induced Pluripotent Stem Cells matrigel Plasmids RNA, Messenger Transforming Growth Factor beta Vitronectin Y 27632
2
Neural Induction of Pluripotent Stem Cells
For dissociating intact colonies of pluripotent stem cells from the layer of DR4 feeders, hiPSCs were exposed to a low concentration of dispase (Invitrogen: 17105-041; 0.7 mg/ml) for ~30 min. Suspended colonies were subsequently transferred into ultra-low-attachment 100 mm plastic plates (Corning) in hiPSC medium without FGF2. For the first 24 h (day 0), the medium was supplemented with the ROCK inhibitor Y-27632 (EMD Chemicals). For neural induction, dorsomorphin (also known as compound C; Sigma 10 μM) and SB-431542 (Tocris, 10 μM) were added to the medium for the first five days. On the sixth day in suspension, the floating spheroids were moved to neural medium (NM) containing Neurobasal (Invitrogen: 10888), B-27 serum substitute without vitamin A (Invitrogen: 12587), GlutaMax (Invitrogen, 1:100), 100 U/ml penicillin and 100 μl streptomycin (Invitrogen). The NM was supplemented with 20 ng/ml FGF2 (R&D Systems) and 20 ng/ml EGF (R&D Systems) for 19 days with daily medium change in the first 10 days, and every other day for the subsequent 9 days. To promote differentiation of the neural progenitors into neurons, FGF2 and EGF were replaced with 20 ng/ml BDNF (Peprotech) and 20 ng/ml NT3 (Peprotech) starting at day 25, while from day 43 onwards only NM without growth factors was used for medium changes every four days.
Paşca A.M., Sloan S.A., Clarke L.E., Tian Y., Makinson C.D., Huber N., Kim C.H., Park J.Y., O’Rourke N.A., Nguyen K.D., Smith S.J., Huguenard J.R., Geschwind D.H., Barres B.A, & Paşca S.P. (2015). Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture. Nature methods, 12(7), 671-678.
Publication 2015
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide dispase dorsomorphin Feeder Cell Layers Fibroblast Growth Factor 2 Growth Factor Human Induced Pluripotent Stem Cells Nervousness Neurons Penicillins Pluripotent Stem Cells Serum Streptomycin Vitamin A Y 27632
3
Directed Differentiation of hPSCs into SC-β Cells

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Publication 2014
Betacellulin Cell Lines Cells Chir 99021 Differentiations, Cell Human Embryonic Stem Cells Human Induced Pluripotent Stem Cells Humidity LDN 193189 Population Group Stem Cells Y 27632
4
Episomal Reprogramming of Fibroblasts to iPSCs
Episomal plasmids and methods were described previously15 (link). Plasmid combination #19 (pEP4-E-O2S-E-T2K, pEP4-E-O2S-E-N2K and pCEP4M2L) was used for most reprogramming unless mentioned otherwise. Plasmids and EBNA mRNA were electroporated into fibroblast cells on Amaxa apparatus according to company instructions. One million cells were used in each electroporation, which were then plated into two 6-well plates. E8 + hydrocortisone media were used for the first 5–10 days, according to cell survival and proliferation after electroporation. When confluency was reached ~20%, hydrocortisone was removed. ES-like iPS cell colonies usually appear after ~25 days. Cells were then picked into individual wells with E8 (TGFβ or NODAL). Cells were passaged for ~ 15 passages before subcloning with Y27632 on Matrigel or vitronectin.
Chen G., Gulbranson D.R., Hou Z., Bolin J.M., Ruotti V., Probasco M.D., Smuga-Otto K., Howden S.E., Diol N.R., Propson N.E., Wagner R., Lee G.O., Antosiewicz-Bourget J., Teng J.M, & Thomson J.A. (2011). Chemically defined conditions for human iPS cell derivation and culture. Nature methods, 8(5), 424-429.
Publication 2011
Cells Cell Survival Electroporation Episomes Fibroblasts Hydrocortisone Induced Pluripotent Stem Cells matrigel Plasmids RNA, Messenger Transforming Growth Factor beta Vitronectin Y 27632
5
Transwell Culture of Human Intestinal Spheroids
Transwell culture methods were adapted from our recently published method for mouse colonic spheroids[19 ]. Human spheroids (~1 well of a 24-well plate per Transwell) were dissociated, strained through a 40-μm filter, seeded onto Transwell membranes (Fisher Scientific, CoStar 3470) coated with 0.1% gelatin (earlier experiments) or Matrigel diluted 1:40 in PBS (later experiments) and provided 5% L-WRN CM (10 μM Y-27632 was included O/N and then removed during daily media changes). TER measurements[19 ] and mucus layer analyses[21 (link)] were performed as previously described. Z-stack images (1.1-μm, with an optimal interval of 0.55-μm) were generated with a Zeiss LSM510 Meta laser scanning confocal microscope (Carl Zeiss Inc., Thornwood, NY) equipped with Argon (Ex. 488 Em. BP 505–530) and HeNe1 (Ex. 543 Em. BP 560–615) lasers, a 63X, 1.4 numerical aperture Zeiss Plan Apochromat oil objective and LSM software. Rectal and ileal spheroid lines were infected with recombinant lentiviruses expressing an enhanced green fluorescent protein (EGFP) under the hPGK promoter [7 (link), 8 (link)] using a described protocol[8 (link)].
VanDussen K.L., Marinshaw J.M., Shaikh N., Miyoshi H., Moon C., Tarr P.I., Ciorba M.A, & Stappenbeck T.S. (2014). Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays. Gut, 64(6), 911-920.
Publication 2014
Argon Colon Culture Techniques enhanced green fluorescent protein Gelatins Homo sapiens Ileum Lentivirus matrigel Microscopy, Confocal Mucus Mus Rectum Tissue, Membrane Y 27632

Most recents protocols related to «Y 27632»

1
Biospecimen Collection for Cancer Studies
Blood samples will be collected before surgery as part of regular medical care. No blood draw will be done specifically for this study. An additional volume of blood will be collected in two 5 ml serum separator tubes and seven 5 ml EDTA tubes and transported to the laboratories to be processed.
Tumor samples will be collected during surgery as part of regular medical care and sent to the pathology laboratory. The surplus tissue (including tumor and adjacent normal tissues) not required for diagnosis will be transported to the laboratory in vials containing medium supplemented in antibiotics and ROCK inhibitor Y-27632.
Perréard M., Florent R., Divoux J., Grellard J.M., Lequesne J., Briand M., Clarisse B., Rousseau N., Lebreton E., Dubois B., Harter V., Lasne-Cardon A., Drouet J., Johnson A., Le Page A.L., Bazille C., Jeanne C., Figeac M., Goardon N., Vaur D., Micault E., Humbert M., Thariat J., Babin E., Poulain L., Weiswald L.B, & Bastit V. (2023). ORGAVADS: establishment of tumor organoids from head and neck squamous cell carcinoma to assess their response to innovative therapies. BMC Cancer, 23, 223.
Full text: Click here
Publication 2023
Antibiotics, Antitubercular BLOOD Blood Volume Diagnosis Edetic Acid Neoplasms Operative Surgical Procedures Serum Tissues Y 27632
2
Establishment and Characterization of PDTO
Tumor pieces will be dissociated enzymatically and mechanically to obtain isolated cells or small cell clusters (Fig. 2). Cells will be embedded in an extracellular matrix (growth factor-reduced Matrigel or BME II) and cultured in a medium supplemented with growth factors and signal pathway inhibitors [Advanced DMEM (Gibco) supplemented with 100 UI/mL of penicillin and streptomycin (Gibco), 1% GlutaMAX (Gibco), 1X B27 (Gibco), 1.25 mM NAC (Sigma-Aldrich), 50 ng/mL EGF (PeproTech), 10 ng/mL FGF-10 (PeproTech), 5 ng/mL FGF-b (PeproTech), 500 nM A-83-01 (PeproTech), 10 μM Y27632 (Interchim), 10 mM Nicotinamide (Sigma-Aldrich), 1 μM PGE2 (PeproTech), 1 μM Forskolin (Peprotech), 0.3 μM CHIR99021 (Biogems), 100 μg/mL Primocin (InvivoGen), 50% Wnt3a, RSPO3, Noggin-conditioned media (L-WRN, ATCC), and 10% RSPO1-conditioned media (Cultrex HA-R-Spondin-1-Fc 293 T, Amsbio)]. Culture medium will be changed twice a week. Once formed, PDTO will be dissociated and reseeded to amplify them for experimental purposes. Cryovials will be prepared at regular intervals by dissociating and resuspending PDTO in Recovery Cell Culture Freezing Medium (Gibco) prior to be biobanked in liquid nitrogen. It should be noted that PDTO line will be considered as established when it will be maintained for more than 3 passages. For each PDTO line, samples will be kept frozen for DNA/RNA/protein analysis and others will be embedded in paraffin for histopathological analysis. This will allow comparisons between the characteristics of the PDTO and the tumor from which they are derived in order to validate their correspondence.

Establishment and characterization of PDTO derived from HNSCC and evaluation of response to treatments to assess its predictive value

Perréard M., Florent R., Divoux J., Grellard J.M., Lequesne J., Briand M., Clarisse B., Rousseau N., Lebreton E., Dubois B., Harter V., Lasne-Cardon A., Drouet J., Johnson A., Le Page A.L., Bazille C., Jeanne C., Figeac M., Goardon N., Vaur D., Micault E., Humbert M., Thariat J., Babin E., Poulain L., Weiswald L.B, & Bastit V. (2023). ORGAVADS: establishment of tumor organoids from head and neck squamous cell carcinoma to assess their response to innovative therapies. BMC Cancer, 23, 223.
Full text: Click here
Publication 2023
Cell Culture Techniques Cells Chir 99021 Colforsin Culture Media Culture Media, Conditioned Dinoprostone Extracellular Matrix Freezing Growth Factor HSP40 Heat-Shock Proteins inhibitors matrigel Neoplasms Niacinamide Nitrogen noggin protein Paraffin Embedding Penicillins Signal Pathways Squamous Cell Carcinoma of the Head and Neck Streptomycin Y 27632
3
Evaluating PDTO Responses to Diverse Treatments
The evaluation of the response to treatments will be performed when PDTO have reached a diameter of 150 μm in « PDTO treatment medium », corresponding to the PDTO culture medium lacking N-Acetylcysteine, Y-27632 and primocin.
PDTO will be collected, resuspended in 2% extracellular matrix/PDTO culture medium and then platted in white and clear bottom 96-well plates previously coated with a 1:1 volume mix of PDTO treatment medium with extracellular matrix. In the case of evaluation of the response to radiotherapy, PDTO will be before irradiated using the CellRad System (FAXITRON Bioptics). In the case of evaluation of the response to chemotherapy or PARP inhibitors, drugs are prepared in 2% extracellular matrix/PDTO culture medium and added 1 hour after PDTO have been plated.
In the case of evaluation of the response to immunotherapies, PDTO will be co-cultured with PDTO specific T cells previously generated (see co-culture of PDTO with immune cells) at a 5:1 ratio. Treatments (such as Nivolumab or Pembrolizumab) will be added directly in the co-culture. A condition containing an MHC-I blocking antibody will be added to control for antigen specific killing.
PDTO morphology will be monitored by taking images during the required time using Incucyte S3 (Sartorius). At the endpoint, PDTO response will be assessed using CellTiter-Glo 3D cell viability assay (Promega) according to the manufacturer’s instruction and luminescence will be measured using GloMax Discover GM3000 (Promega) with the associated software. Results will be normalized to the control condition. IC50 will be calculated with GraphPad software. The ability of T cells to recognize and induce lysis of PDTO will be monitored via analysis of caspase 3 cleavage within PDTO and visualization of LAMP-1 on the membrane of CD8+ T cells.
The treatment response of the PDTO will be finally compared to the clinical response (PFS/DFS/OS) of the patient from whom they are derived in order to validate the predictive value of this model for HNSCC.
Perréard M., Florent R., Divoux J., Grellard J.M., Lequesne J., Briand M., Clarisse B., Rousseau N., Lebreton E., Dubois B., Harter V., Lasne-Cardon A., Drouet J., Johnson A., Le Page A.L., Bazille C., Jeanne C., Figeac M., Goardon N., Vaur D., Micault E., Humbert M., Thariat J., Babin E., Poulain L., Weiswald L.B, & Bastit V. (2023). ORGAVADS: establishment of tumor organoids from head and neck squamous cell carcinoma to assess their response to innovative therapies. BMC Cancer, 23, 223.
Full text: Click here
Publication 2023
Acetylcysteine Antibodies, Blocking Antigens Biological Assay Caspase 3 CD8-Positive T-Lymphocytes Cells Cell Survival Coculture Techniques Cultured Cells Culture Media Cytokinesis Extracellular Matrix Immunotherapy Luminescence lysosomal-associated membrane protein 1, human Nivolumab Patients pembrolizumab Pharmaceutical Preparations Pharmacotherapy Poly(ADP-ribose) Polymerase Inhibitors Promega Squamous Cell Carcinoma of the Head and Neck T-Lymphocyte Tissue, Membrane Y 27632
4
Immunofluorescent Characterization of iPSC-Derived Cardiomyocytes
IPSc-derived cardiomyocytes were cultured on glass slides coated with Synthemax II-SC (Corning) in RPMI-1640/B-27 with Y27632 (5 μM) media for 2 days. Cells were fixed with 4% formaldehyde for 15 min at room temperature. Fixed cells were blocked in antibody buffer (5% BSA, 0.1% Tween-20 in PBS) for 1 h at room temperature. Following blocking, cells were incubated overnight at 4 °C with cardiac troponin-T antibody (abcam, ab45932, 1:200), α-Actinin antibody (Sigma, A7811, 1:800), and/or Connexin 43 (Cell Signaling Technology, 1:100) in antibody buffer. After the overnight incubation, cells were washed three times in antibody buffer. Following washing, cells were incubated with Alexa-Fluor conjugated secondary antibodies (Thermo Fisher Scientific, Alexa 488 Donkey anti-Rabbit IgG and Alexa 594 Goat anti-Ms IgG1) at a 1:1,000 dilution in antibody buffer for 1 h at room temperature. Nuclei were stained by DAPI at 1 μg/ml and Wheat Germ Agglutinin (Thermo Fisher Scientific, W21404) was used to stain cell membrane for 10 min at room temperature in antibody buffer. Following washing in PBS to remove unbound complexes, sarcomeres were analyzed using a Zeiss LSM510 confocal microscope. Images were processed with Zeiss software (Axiovision Rel4.8 and Zen Blue). Circularity measurements were made by comparing cardiomyocyte length to width and were expressed as a circularity index whereby circularity index = width/length15 (link). Sarcomere distance measurements were made with ImageJ15 (link). All measurements were made with a double-blinding method.
Hsueh Y.C., Pratt R.E., Dzau V.J, & Hodgkinson C.P. (2023). Novel method of differentiating human induced pluripotent stem cells to mature cardiomyocytes via Sfrp2. Scientific Reports, 13, 3920.
Full text: Click here
Publication 2023
Actinin Alexa594 anti-IgG Antibodies Buffers Cardiac Arrest Cell Nucleus DAPI Equus asinus Formaldehyde GJA1 protein, human Goat Heart IgG1 Immunoglobulins Induced Pluripotent Stem Cells Microscopy, Confocal Myocytes, Cardiac Plasma Membrane Rabbits Sarcomeres Stains Technique, Dilution Troponin T Tween 20 Wheat Germ Agglutinins Y 27632
5
Electrophysiological Recordings of iPSC-Derived Cardiomyocytes
IPSc-derived cardiomyocytes were cultured on glass slides coated with BG iMatrix-511 (Biogems, 0.5 μg/cm2) in RPMI-1640/B-27 with Y27632 (5 μM) media for 2 days. The microelectrodes (Sutter Instruments, BF150-110-10) were made via a micropipette puller (Sutter Instrument, P-87) to create electrodes at ~ 3 MΩ in Tyrode's solution (140 mM NaC1, 5.4 mM KCl, 1.8 mM CaCl2, 1.05 mM MgCl2, 0.33 mM NaH2PO4, 5 mM HEPES and 10 mM glucose; pH was adjusted to 7.4 with NaOH). Current-clamp mode was used to record action potential via an Axopatch 200B amplifier running software Calmpex 8.2.
Hsueh Y.C., Pratt R.E., Dzau V.J, & Hodgkinson C.P. (2023). Novel method of differentiating human induced pluripotent stem cells to mature cardiomyocytes via Sfrp2. Scientific Reports, 13, 3920.
Full text: Click here
Publication 2023
Action Potentials Glucose HEPES Induced Pluripotent Stem Cells Magnesium Chloride Microelectrodes Myocytes, Cardiac Tyrode's solution Y 27632

Top products related to «Y 27632»

1
Y 27632 by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, Canada, Australia, Switzerland, Israel, China, Morocco
Y-27632 is a selective and potent Rho-associated protein kinase (ROCK) inhibitor. It functions by inhibiting the activity of ROCK, a key enzyme involved in various cellular processes.
2
Matrigel by Corning
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Matrigel is a complex mixture of extracellular matrix proteins derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is widely used as a basement membrane matrix to support the growth, differentiation, and morphogenesis of various cell types in cell culture applications.
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Y 27632 by Bio-Techne
Sourced in United Kingdom, United States, Canada
Y-27632 is a Rho-associated protein kinase (ROCK) inhibitor. It functions by selectively inhibiting the activity of ROCK, a key regulator of actin cytoskeleton organization and cell contractility.
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Glutamax by Thermo Fisher Scientific
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GlutaMAX is a chemically defined, L-glutamine substitute for cell culture media. It is a stable source of L-glutamine that does not degrade over time like L-glutamine. GlutaMAX helps maintain consistent cell growth and performance in cell culture applications.
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Y 27632 by Selleck Chemicals
Sourced in United States, Germany, China, United Kingdom, Canada
Y-27632 is a cell-permeable, selective and potent inhibitor of Rho-associated protein kinase (ROCK). It functions by inhibiting ROCK activity, which plays a crucial role in regulating cellular processes such as cytoskeleton organization, cell migration, and cell proliferation.
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Dmem f12 by Thermo Fisher Scientific
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DMEM/F12 is a cell culture medium developed by Thermo Fisher Scientific. It is a balanced salt solution that provides nutrients and growth factors essential for the cultivation of a variety of cell types, including adherent and suspension cells. The medium is formulated to support the proliferation and maintenance of cells in vitro.
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Penicillin streptomycin by Thermo Fisher Scientific
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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Y 27632 by Fujifilm
Sourced in Japan, United States
Y-27632 is a pharmacological agent that inhibits the activity of Rho-associated protein kinase (ROCK). It is a widely used tool in cell biology research for its ability to modulate cell contractility and cytoskeletal organization.
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Y 27632 by STEMCELL
Sourced in Canada, United States, Germany, United Kingdom
Y-27632 is a small molecule Rho-associated protein kinase (ROCK) inhibitor. It is commonly used in cell culture applications to promote the survival and growth of various cell types.
10
Matrigel by BD
Sourced in United States, United Kingdom, Germany, China, Canada, Japan, Italy, France, Belgium, Australia, Uruguay, Switzerland, Israel, India, Spain, Denmark, Morocco, Austria, Brazil, Ireland, Netherlands, Montenegro, Poland
Matrigel is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, a tumor rich in extracellular matrix proteins. It is widely used as a substrate for the in vitro cultivation of cells, particularly those that require a more physiologically relevant microenvironment for growth and differentiation.

More about "Y 27632"

Y-27632 is a Rho-associated protein kinase (ROCK) inhibitor that has been extensively studied for its potential therapeutic applications.
This compound has demonstrated the ability to modulate various cellular processes, including cell migration, proliferation, and differentiation.
Researchers utilize Y-27632 to investigate its effects on a range of biological systems and disease models, such as neurological disorders, cardiovascular diseases, and cancer.
When conducting experiments with Y-27632, researchers often use additional reagents and materials, such as Matrigel, a basement membrane matrix used for cell culture and 3D assays; GlutaMAX, a glutamine supplement for cell culture; and DMEM/F12, a cell culture medium that provides essential nutrients.
Additionally, Penicillin/streptomycin, an antibiotic mixture, is commonly used to prevent bacterial contamination in cell culture experiments.
The detailed description and comparison of Y-27632 research protocols can help optimize experiments and lead to more informed decisions, streamlining the discovery process.
PubCompare.ai's AI-driven tools can assist researchers in locating relevant protocols from literature, preprints, and patents, and identify the best options using advanced comparisons.
By utilizing these resources, researchers can enhance their Y-27632 studies and make more informed decisions, ultimately accelerating the discovery and development of potential therapies.