High-content screening of small compounds on human embryonic stem cells.

Human ES (embryonic stem) cells and iPS (induced pluripotent stem) cells have been heralded as a source of differentiated cells that could be used in the treatment of degenerative diseases, such as Parkinson's disease or diabetes. Despite the great potential for their use in regenerative therapy, the challenge remains to understand the basic biology of these remarkable cells, in order to differentiate them into any functional cell type. Given the scale of the task, high-throughput screening of agents and culture conditions offers one way to accelerate these studies. The screening of small-compound libraries is particularly amenable to such high-throughput methods. Coupled with high-content screening technology that enables simultaneous assessment of multiple cellular features in an automated and quantitative way, this approach is proving powerful in identifying both small molecules as tools for manipulating stem cell fates and novel mechanisms of differentiation not previously associated with stem cell biology. Such screens performed on human ES cells also demonstrate the usefulness of human ES/iPS cells as cellular models for pharmacological testing of drug efficacy and toxicity, possibly a more imminent use of these cells than in regenerative medicine.

[1]  Albert Gough,et al.  High-Content Screening: A New Approach to Easing Key Bottlenecks in the Drug Discovery Process , 1997 .

[2]  J. Miyazaki,et al.  Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells , 2000, Nature Genetics.

[3]  Hynek Wichterle,et al.  Induced Pluripotent Stem Cells Generated from Patients with ALS Can Be Differentiated into Motor Neurons , 2008, Science.

[4]  D. Rancourt,et al.  ROCK inhibitor improves survival of cryopreserved serum/feeder-free single human embryonic stem cells. , 2008, Human reproduction.

[5]  S. Schreiber,et al.  Small molecules efficiently direct endodermal differentiation of mouse and human embryonic stem cells. , 2009, Cell stem cell.

[6]  T. Mitchison,et al.  Towards a pharmacological genetics. , 1994, Chemistry & biology.

[7]  Sheng Ding,et al.  Generation of rat and human induced pluripotent stem cells by combining genetic reprogramming and chemical inhibitors. , 2009, Cell stem cell.

[8]  P. Schultz,et al.  A small molecule primes embryonic stem cells for differentiation. , 2009, Cell stem cell.

[9]  Peter G Schultz,et al.  Small molecules that induce cardiomyogenesis in embryonic stem cells. , 2004, Journal of the American Chemical Society.

[10]  J. Leikola Society Transactions , 1976 .

[11]  J. Comley,et al.  A 1536 Colorimetric SPAP Reporter Assay: Comparison with 96- and 384-Well Formats , 1998 .

[12]  Robert Nadon,et al.  Statistical practice in high-throughput screening data analysis , 2006, Nature Biotechnology.

[13]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[14]  S. Strickland,et al.  The induction of differentiation in teratocarcinoma stem cells by retinoic acid , 1978, Cell.

[15]  Thomas D. Y. Chung,et al.  A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays , 1999, Journal of biomolecular screening.

[16]  Ivana Barbaric,et al.  Appearances can be deceiving: phenotypes of knockout mice. , 2007, Briefings in functional genomics & proteomics.

[17]  P. Andrews,et al.  Specific Knockdown of Oct4 and β2‐microglobulin Expression by RNA Interference in Human Embryonic Stem Cells and Embryonic Carcinoma Cells , 2004, Stem cells.

[18]  Lani F. Wu,et al.  Image-based multivariate profiling of drug responses from single cells , 2007, Nature Methods.

[19]  P. Andrews Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. , 1984, Developmental biology.

[20]  P. Cohen,et al.  The selectivity of protein kinase inhibitors: a further update. , 2007, The Biochemical journal.

[21]  J. Stelling,et al.  Robustness of Cellular Functions , 2004, Cell.

[22]  James A. Thomson,et al.  Homologous recombination in human embryonic stem cells , 2003, Nature Biotechnology.

[23]  P. Cohen,et al.  Specificity and mechanism of action of some commonly used protein kinase inhibitors. , 2000, The Biochemical journal.

[24]  Wenjun Guo,et al.  Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2 , 2008, Nature Biotechnology.

[25]  B. Thiers Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2008 .

[26]  N. Socci,et al.  BAC Transgenesis in Human Embryonic Stem Cells as a Novel Tool to Define the Human Neural Lineage , 2009, Stem cells.

[27]  Marius Wernig,et al.  In vitro differentiation of transplantable neural precursors from human embryonic stem cells , 2001, Nature Biotechnology.

[28]  Sheng Ding,et al.  Induction of pluripotent stem cells from mouse embryonic fibroblasts by Oct4 and Klf4 with small-molecule compounds. , 2008, Cell stem cell.

[29]  N. Socci,et al.  High-throughput screening assay for the identification of compounds regulating self-renewal and differentiation in human embryonic stem cells. , 2008, Cell stem cell.

[30]  C. Peterson,et al.  Stem cell states, fates, and the rules of attraction. , 2009, Cell stem cell.

[31]  George Q. Daley,et al.  Disease-Specific Induced Pluripotent Stem Cells , 2008, Cell.

[32]  Gordon Keller,et al.  Differentiation of Embryonic Stem Cells to Clinically Relevant Populations: Lessons from Embryonic Development , 2008, Cell.

[33]  P. Andrews,et al.  Novel regulators of stem cell fates identified by a multivariate phenotype screen of small compounds on human embryonic stem cell colonies. , 2010, Stem cell research.

[34]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[35]  Peter G Schultz,et al.  Reprogramming of murine fibroblasts to induced pluripotent stem cells with chemical complementation of Klf4 , 2009, Proceedings of the National Academy of Sciences.

[36]  H. Schöler,et al.  Self-renewal of embryonic stem cells by a small molecule , 2006, Proceedings of the National Academy of Sciences.

[37]  S. Schreiber Chemical genetics resulting from a passion for synthetic organic chemistry. , 1998, Bioorganic & medicinal chemistry.

[38]  S. Nishikawa,et al.  A ROCK inhibitor permits survival of dissociated human embryonic stem cells , 2007, Nature Biotechnology.

[39]  W. Scher,et al.  Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Y. Kawazoe,et al.  [Synthetic small molecules that control stem cell fate]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[41]  A. Viale,et al.  Modeling Pathogenesis and Treatment of Familial Dysautonomia using Patient Specific iPSCs , 2009, Nature.