hiPS Derived Cardiomyocyte Assay for Screening Toxicant Differentiation Effects

用于筛选毒性分化效应的 hiPS 衍生心肌细胞测定

• 批准号: 8618274
• 负责人: Bradley H. Garcia
• 金额: $ 21.08万
• 依托单位: PRIMORIGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8618274
• 关键词: Address; Affect; Animal Model; Biological Assay; Brachyury protein; Calcium; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cell Line; Cell Survival; Cells; Collection; Data; Development; Developmental Process; Dimethyl Sulfoxide; Ethidium; Evaluation; Flow Cytometry; Fluorescence; Funding; Genetic Variation; Government; Growth Factor; Heart Atrium; Heart Diseases; Hip region structure; Human; Human Cell Line; Image Analysis; Late Effects; Libraries; Measurement; Mesoderm; Methods; Monitor; National Institute of Environmental Health Sciences; Nodal; Optics; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Population; Population Heterogeneity; Preclinical Drug Evaluation; Predictive Value; Prevalence; Process; Protocols documentation; Research; Risk; Safety; Services; Small Business Innovation Research Grant; Source; Staging; Staining method; Stains; Stem cells; System; Technology; Teratogens; Testing; Thalidomide; Toxic effect; Toxicology; Toxin; Troponin I; Troponin T; Ventricular; Video Microscopy; Work; base; calcein AM; cell type; cytokine; cytotoxicity; high throughput screening; human tissue; induced pluripotent stem cell; member; novel; phase 1 study; phase 2 study; programs; public health relevance; scale up; screening; small molecule; toxicant; toxicant screening

Primorigen Biosciences (R) Abstract Primorigen Biosciences will use SBIR funds to develop a high-throughput toxicology assay system utilizing human ventricular cardiomyocytes (vCMs) differentiated from human induced pluripotent stem cells (hiPSCs). Phase I studies will convert the small-molecule based, cytokine-free CardioTotal" differentiation technology into a 96-well based high-throughput format. Automated flow cytometry will be used to monitor mesoderm specification by staining for brachyury, and to monitor differentiation efficiency and yield by staining for cardiac Troponin T and cardiac Troponin I. Cell viability and yield will be determined by cytoxicity assay, and contractility measurements will be performed by video microscopy and automated image analysis. Using these assays, a library of 20 toxicants selected from the Tox21 library will be screened, first for effects upon differentiation following early exposure, and second for toxicity and contractility effects by late exposure of differentiated cardiomyocytes. In Phase II, the system will be scaled up for 384-well format, and screening will be applied to a larger portion of the Tox21 library. Additional pluripotent cell lines will be tested to establish robustness, and secondary screenings will be initiated to identify mechanisms of action. In Phase III, the system will be commercialized as an optimized, high-throughput cardiotoxicity screening system, provided as a service by Primorigen or customized for end-user internalization (for example at NIEHS NTP or EPA's ToxCast program).

Primorigen Biosciences (R)摘要