TR&D1: Stem Cell and Induced Pluripotent Stem Cell Resources (Pages 116-134)

该中心致力于开发用于鉴定和表征与细胞糖蛋白和糖脂相关的聚糖结构和转录物的技术和工具。作为进一步开发糖组学技术和工具的试验平台，研究将再次关注人类多能干细胞的特征，包括胚胎干细胞（hESC）和诱导多能干细胞（iPSC）。一系列来源于干细胞的分化细胞谱系将纳入本分析。这种生物模型是糖组学技术和工具开发的理想试验平台，因为它提供了各种各样的聚糖结构和挑战，这是由于可以产生的细胞类型的多样性。这些研究将为人类胚胎发育提供新的见解，并将导致识别用于干细胞表征的新生物标志物。该TR&D的主要优势在于其成功地从hESC和hiPSC中产生了纯的、充分表征的细胞类型。例如，我们常规生产适合下游分析的神经、肝脏、胰腺和心脏谱系。第一种从多能细胞直接分化神经嵴干细胞的方法[1]是作为该中心上一轮资金的一部分开发的。另外的心血管谱系也正在开发中，以增加其他TR& D分析的细胞的多样性，并提高将遇到的聚糖复杂性。 在这次竞争性更新中，我们建议扩展我们以前的工作，并将新的重点放在糖基化相关疾病的人类iPSC模型上。了解这些疾病的基础是一项重大挑战，也是一个基本上探索不足和了解不多的领域。使用iPSC重编程技术，我们建议生成一组完全验证的细胞系，可以作为人类先天性糖基化疾病（CDG）和先天性肌营养不良症（CMD）的模型。这将为大量研究人员提供宝贵的资源，并为糖组学技术开发创造一个真正具有挑战性的试验平台。该TR&D的另一个主要新组成部分是将Richard Steet博士列为共同研究者。Steet博士将把他在先天性糖基化疾病领域的广泛专业知识带到这项工作中，并将主要负责所得细胞系的生化验证。 博士Steet还将协调本TR&D中许多DBP合作者和人员之间的互动。

This Center is dedicated to the development of technologies and tools for the identification and characterization of glycan structures and transcripts associated with cellular glycoproteins and glycolipids. As a test-bed for the further development of glycomic technologies and tools, studies will again focus on the characterization of human pluripotent stem cells, including embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). A range of differentiated cell lineages derived from stem cells will be incorporated into this analysis. This biological model is ideal as a test-bed for glycomics technologies and tool development because it offers a diverse range of glycan structures and challenges due to the diversity in cell types that can be generated. These studies will offer new insights into human embryonic development and will lead to the identification of new biomarkers for stem cell characterization. A major strength of this TR&D has been its success in generating pure, well-characterized cell types from hESC and hiPSCs. For example, we routinely produce neural, liver, pancreatic and cardiac lineages that are suitable for downstream analysis. The first method for direct differentiation of neural crest stem cells from pluripotent cells [1] was developed as part of the Center's previous round of funding. Additional cardiovascular lineages are also being developed to increase the diversity of cells to be analyzed by other TR&Ds'and to enhance glycan complexity that will be encountered. In this competitive renewal, we propose to extend our previous work and also include a new emphasis on human iPSC models for glycosylation-related disease. Understanding the basis of these diseases represents a major challenge and an area that is largely underexplored and poorly understood. Using iPSC-reprogramming technology, we propose to generate panels of fully validated cell lines that can serve as models for human congenital disorders of glycosylation (CDG) and congenital muscular dystrophy (CMD). This will serve to provide valuable resources for a large number of investigators and to create a truly challenging test-bed for glycomics technology development. Another major new component of this TR&D is the inclusion of Dr. Richard Steet as co-investigator. Dr. Steet will bring his broad expertise in the area of congenital glycosylation disorders to this effort and will be primarily responsible for the biochemical validation of the resulting cell lines. Dr. Steet will also coordinate the interactions between many DBP collaborators and personnel within this TR&D.