Development of Enabling Suspension Culture & Drug Screening Technologies

人胚胎干细胞（hESC）是一种非常有前途的分化细胞来源， 在未来，用于糖尿病、帕金森氏症和其他疾病的革命性治疗。hESC已经被 研究起来有些困难，因为支持它们在文化中生长的条件非常原始。我们 已经开发了一种简单有效的方法来培养hESC，这开辟了替代和 强大的细胞培养方法，这可能会加速这些临床应用的进展。 该申请的目的是为hESC开发关键的新技术。我们有 建立了一种新的、简便的hESC成分确定培养基体系。我们的方法 支持未分化细胞的大量扩增，维持悬浮的hESC， hESC培养物的常规小型化用于高通量筛选方法。具体目标是 研究将未分化的hESC保持在悬浮液中并指导分化的优点， 内胚层和其他谱系在高密度悬浮培养。将调整简化的方案 到完全可控的生物反应器系统并优化。其他目的是筛选已知的化合物或 hESC中的海洋天然产物的文库，以鉴定影响自我更新的新活性， 分化或凋亡。特别地，我们将通过以下方法筛选诱导hESC分化的化合物： 影响激活素/nodal、GSKSp或PIS激酶信号传导。我们的方法非常独特， 极大地简化hESC衍生的可移植细胞的未来细胞治疗应用的潜力。

Human embryonic stem cells (hESCs) are a highly promising source of differentiated cells that may be used in the future for revolutionary treatments of diabetes, Parkinson's and other diseases. hESCs have been somewhat difficult to study, because conditions that support their growth in culture were farily primitive. We have developed a simple and efficient way to culture hESCs, which opens the possibility of alternate and powerful cell culture methodologies, which may speed the progress of these clinical applications. The ojectives of the application are to develop key new enabling technologies for hESCs. We have established a new and simple defined media system for culturing and differentiating hESCs. Our approach supported the massive expansion of undifferenitiated cells, the maintenance of hESCs in suspension and the routine minaturization of hESC culture for high throughtput sceening approaches. The specific aims are to investigate the advantages of maintaining undifferenitated hESCs in suspension, and directing differentiation to endodermal and other lineages in high-density suspension culture. Streamlined protocols will be adapted to a fully-controlable bioreactor system and optimised. Other aims are to screen known compounds or libaries of marine natural products in hESCs to identify novel activities that impact self-renewal, differentiation or apoptosis. In particular, we will screen for compunds that induce differenitation in hESCs by impacting Activin/nodal, GSKSp or PIS Kinase signaling. Our approaches are highly unique and have the potential to greatly simplify future cell therapy applications of hESC-derived transplantable cells.