New hiPSC tools for astroglial-focused approaches to ALS

新的 hiPSC 工具用于以星形胶质细胞为中心的 ALS 治疗方法

• 批准号: 8293969
• 负责人: KEVIN C EGGAN
• 金额: $ 84.96万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8293969
• 关键词: Activities of Daily Living; Acute; Affinity; Amyotrophic Lateral Sclerosis; Antibodies; Astrocytes; Binding Proteins; Biochemical; Biological Assay; Cell Differentiation process; Cell Line; Cell Separation; Cell Transplantation; Cells; Cervical; Connexin 43; Coupling; Cues; Development; Electrophysiology (science); Embryo; Engraftment; Evaluation; Excitatory Amino Acid Transporter 2; Exhibits; FK506; Family; Fingers; Fluo 4; Fluorescence; Future; GLAST Protein; Generations; Genes; Genome; Glial Fibrillary Acidic Protein; Glutamate Transporter; Glutamates; Grant; Gray unit of radiation dose; Green Fluorescent Proteins; Human; Image; Immunohistochemistry; Immunosuppression; In Vitro; Individual; Injection of therapeutic agent; Insertional Mutagenesis; Knock-in Mouse; Laboratories; Lactate Transporter; Life; Mediating; Membrane Potentials; Methods; Monitor; Morphology; Mutation; Neurons; Phenotype; Physiology; Potassium; Preclinical Drug Evaluation; Property; Proteins; Protocols documentation; Purinoceptor; Reporter; Reporter Genes; Rest; Rodent; Schwann Cells; Screening procedure; Signal Transduction; Site; Slice; Sorting - Cell Movement; Specificity; Spinal Cord; Stem cells; Technology; Time; Transplantation; Whole-Cell Recordings; Work; Zinc Fingers; aldehyde dehydrogenases; aquaporin 4; base; disease phenotype; enhanced green fluorescent protein; experience; follow-up; homologous recombination; in vivo; induced pluripotent stem cell; injured; juvenile animal; nuclease; promoter; protein aggregate; protein expression; research study; response; tool; transcription factor; uptake; water channel

At the time of submission of the orginal ALS iPS Go Grant, there were no well-validated protocols for generating astroglia from human ES or IPS cell lines, although approaches from rodent cells existed. Fortunately, we were able to develop a reliable protocol in both the Henderson lab and the Rothstein labs (Figure 5A). We now have a protocol that works identically in both labs¿with very similar efficiency at differentiation between labs. We have also been able to detect evidence of true astroglial maturation in a number of lines that have advanced sufficiently and these express appropriate astroglial specific proteins and have functional glutamate transport (Figure 5B-D).

在提交最初的ALS iPS Go Grant时，虽然存在啮齿动物细胞的方法，但还没有从人类胚胎干细胞或iPS细胞系中产生星形胶质细胞的经过良好验证的方案。