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Chemical Structure| 146986-50-7 Chemical Structure| 146986-50-7

Structure of Y-27632
CAS No.: 146986-50-7

Chemical Structure| 146986-50-7

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Y-27632 is an orally active, ATP-competitive inhibitor of ROCK-I and ROCK-II with Ki values of 220 nM and 300 nM, respectively.

Synonyms: Y-27632

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Pierson Smela, Merrick ;

Abstract: The ovary is the central organ of female reproduction, and understanding it is crucial for improving women’s health and treating infertility. However, studies of human ovarian development are limited by technical and ethical challenges in accessing ovarian tissue, especially during fetal stages. Therefore, engineering stem cell-based models of the human ovary in vitro has the potential to rapidly advance the study of human reproduction by enabling experiments that would not otherwise be possible. Furthermore, if the full process of human oogenesis can be re-created in vitro starting from pluripotent stem cells, this would be a transformative therapy for millions of people around the world who are infertile due to a lack of eggs. Although in vitro oogenesis has recently been achieved in mice, this has not yet been successful for humans, largely due to a lack of methods for producing the necessary ovarian cell types. In this dissertation, I present my work on directing the differentiation of human induced pluripotent stem cells (hiPSCs) into ovarian cell typesin vitro, including granulosa-like, oogonia-like, and meiotic cells. My overall approach has centered on using computational tools and multiplexed screens to identify key regulatory factors that drive the formation of a cell type of interest. In Chapter 1, I discuss the production of granulosa-like cells and their application in forming ovarian organoids. In Chapter 2, I showcase a method for the rapid generation of oogonia-like cells with erasure of DNA methylation. In Chapter 3, I describe my latest work on directly initiating meiosis from hiPSCs. Overall, these methods establish in vitro models of various aspects of ovarian development that will be useful to researchers in the field, and may serve as the basis for future therapies such as in vitro oogenesis.

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Pierson Smela, Merrick D. ; Kramme, Christian C. ; Fortuna, Patrick R. J. ; Adams, Jessica L. ; Su, Rui ; Dong, Edward , et al.

Abstract: An in vitro model of human ovarian follicles would greatly benefit the study of female reproduction. Ovarian development requires the combination of germ cells and several types of somatic cells. Among these, granulosa cells play a key role in follicle formation and support for oogenesis. Whereas efficient protocols exist for generating human primordial germ cell-like cells (hPGCLCs) from human induced pluripotent stem cells (hiPSCs), a method of generating granulosa cells has been elusive. Here, we report that simultaneous overexpression of two transcription factors (TFs) can direct the differentiation of hiPSCs to granulosa-like cells. We elucidate the regulatory effects of several granulosa-related TFs and establish that overexpression of NR5A1 and either RUNX1 or RUNX2 is sufficient to generate granulosa-like cells. Our granulosa-like cells have transcriptomes similar to human fetal ovarian cells and recapitulate key ovarian phenotypes including follicle formation and steroidogenesis. When aggregated with hPGCLCs, our cells form ovary-like organoids (ovaroids) and support hPGCLC development from the premigratory to the gonadal stage as measured by induction of DAZL expression. This model system will provide unique opportunities for studying human ovarian biology and may enable the development of therapies for female reproductive health.

Keywords: developmental biology ; granulosa ; human ; mouse ; oogenesis ; ovary ; regenerative medicine ; stem cells ; transcription factors

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Product Details of Y-27632

CAS No. :146986-50-7
Formula : C14H21N3O
M.W : 247.34
SMILES Code : O=C([C@H]1CC[C@H]([C@H](N)C)CC1)NC2=CC=NC=C2
Synonyms :
Y-27632
MDL No. :MFCD03093855
InChI Key :IDDDVXIUIXWAGJ-DDSAHXNVSA-N
Pubchem ID :9901617

Safety of Y-27632

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H312-H332
Precautionary Statements:P280

Related Pathways of Y-27632

cytoskeleton

Isoform Comparison

Biological Activity

In Vitro:

Cell Line
Concentration Treated Time Description References
ARPE19 cells 40µM 24 h Y-27632 treatment significantly increased nanotube formation and mitochondrial transfer rate PMC11426248
ARPE19 cells 10µM 24 h Y-27632 treatment led to an increase in nanotube formation and enhanced mitochondrial transfer among ARPE19 cells PMC11426248

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
New Zealand white rabbits Glaucoma model Topical instillation 2% w/v, 50 μL Single dose, lasting for 10 days To evaluate the therapeutic effect of HMCNs on glaucoma, results showed that HMCNs with a 20 nm shell thickness effectively reduced intraocular pressure and repaired damaged cornea and retina. PMC8039939
Mice OVA sensitised and RSV infected mouse model Oral gavage 30 mg/kg Administered 2 hours before methacholine challenge Y-27632 significantly reduced baseline lung resistance in OVA sensitised mice and suppressed methacholine and serotonin induced airway hyperresponsiveness in both OVA sensitised and OVA/RSV mice PMC1746359

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

4.04mL

0.81mL

0.40mL

20.22mL

4.04mL

2.02mL

40.43mL

8.09mL

4.04mL

References

 

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