Human embryonic stem cell

人类胚胎干细胞

• 批准号: 8557124
• 负责人: Pamela Robey
• 金额: $ 79.36万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8557124
• 关键词: Biochemical Pathway; Cell Culture System; Cell Culture Techniques; Cell Death; Cell Line; Cell Survival; Cells; Cellular Stress; Chromosomal Instability; Clinic; Collaborations; Communities; Controlled Study; DNA; Data; Deposition; Differentiation Antigens; ES Cell Line; Educational process of instructing; Embryo; Evaluation; Gene Expression; Generations; Genes; Genetic; Genome; Genome Stability; Goals; Human; MDM2 gene; Mentors; Methods; Mission; Monitor; Neurons; Pathway interactions; Patients; Pluripotent Stem Cells; Population; Process; Protocols documentation; Quality Control; RNA; Regulation; Reporter; Research Personnel; Role; Sampling; Side; Site; Somatic Cell; Sorting - Cell Movement; Source; Staging; Stem Cell Research; Stem cells; Surface; System; Technology; Testing; United States National Institutes of Health; Update; Xeno; Zinc Fingers; base; biological adaptation to stress; cell growth; human embryonic stem cell; immunocytochemistry; improved; induced pluripotent stem cell; inhibitor/antagonist; insight; interest; monolayer; new technology; novel; novel strategies; progenitor; recombinase; scale up; small molecule; stem cell technology; user-friendly; vector; web site

We have performed extensive characterization of the 21 Bush-era human embryonic stem cells and deposited the data with NCBI GEO for public access. We have created a user-friendly gene expression search engine which allows a casual user to interrogate the data for their particular gene of interest. In our mission to facilitate pluripotent stem cell research, we have performed in-depth characterization of a control induced pluripotent stem cell (iPSC) line, BC1. This includes FACS analysis and immunocytochemistry as well as RNA extraction for gene expression analysis which will be performed on Agilent microarray chips. We have also generated some reporter embryonic stem cell lines which are currently undergoing evaluation and characterization. In terms of bringing pluripotent stem cells to the clinic we have been evaluating novel xeno-free substrates, media and small molecule inhibitors. In addition, since many iPSC lines have been generated on campus using a LoxP flanked polycistronic vector we have also been testing methods to excise the foreign DNA with Cre recombinase. In collaboration with the NIH CRM, we have generated iPSCs derived from neuronal precursors differentiated from H1 ES cell lines for comparison them to the parental line. These isogenic lines will enable a clear comparison of hESC and hiPSCs. We have continued the study of critical genetic and biochemical pathways that control genome stability, cellular stresses, cell growth, and differentiation. We have generated an efficient cell culture system based on the non-colony type monolayer method for controlling pluripotent cell growth. We have also provided important insights into the role of surface markers in the regulation of the pluripotent states and differentiation processes of human pluripotent stem cells. Finally, we have been involved in mentoring and teaching standard and feeder-free, pluripotent stem cell culture, assisting and advising on the generation of iPSCs from patient samples as well as assisting and advising on differentiation strategies as requested. We update the SCU website with protocols and information as it becomes available to aid other researchers in their studies.

我们对 21 个布什时代的人类胚胎干细胞进行了广泛的表征，并将数据存入 NCBI GEO 供公众访问。我们创建了一个用户友好的基因表达搜索引擎，允许临时用户查询他们感兴趣的特定基因的数据。为了促进多能干细胞研究，我们对对照诱导多能干细胞 (iPSC) 系 BC1 进行了深入表征。这包括 FACS 分析和免疫细胞化学以及用于基因表达分析的 RNA 提取，这些分析将在安捷伦微阵列芯片上进行。我们还生成了一些报告胚胎干细胞系，目前正在进行评估和表征。 在将多能干细胞引入临床方面，我们一直在评估新型无异源底物、培养基和小分子抑制剂。此外，由于使用 LoxP 侧翼多顺反子载体在校园内生成了许多 iPSC 系，我们还一直在测试使用 Cre 重组酶切除外源 DNA 的方法。我们与 NIH CRM 合作，生成了源自 H1 ES 细胞系分化的神经元前体的 iPSC，以便将它们与亲本系进行比较。这些同基因系将能够清晰比较 hESC 和 hiPSC。 我们继续研究控制基因组稳定性、细胞应激、细胞生长和分化的关键遗传和生化途径。我们基于非集落型单层方法构建了一种有效的细胞培养系统，用于控制多能细胞的生长。我们还对表面标记在调节人类多能干细胞的多能状态和分化过程中的作用提供了重要的见解。 最后，我们参与指导和教授标准和无饲养层多能干细胞培养，协助和建议从患者样本中生成 iPSC，并根据要求协助和建议分化策略。我们会更新 SCU 网站的协议和信息，以帮助其他研究人员进行研究。