Defining fate potential in human ESC derived neural stem cells

定义人类 ESC 衍生神经干细胞的命运潜力

• 批准号: 7713919
• 负责人: LORENZ P. STUDER
• 金额: $ 51.42万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7713919
• 关键词: Address; Adult; Affect; Anterior; Apical; Ascorbic Acid; Astrocytes; Biological Assay; Biology; Brain; Cell Therapy; Cell physiology; Cells; Cues; Data; Derivation procedure; Development; Dopamine; Engraftment; Epiblast; Epigenetic Process; Erinaceidae; Exhibits; FGF8 gene; Fibroblast Growth Factor 2; Gene Transfer Techniques; Genes; Genetic; Glutamates; Goals; Heterogeneity; Human; In Vitro; Individual; Interneurons; Intrinsic factor; Maintenance; Midbrain structure; Molecular; Motor Neurons; Nature; Nervous system structure; Neural Crest; Neuroepithelial; Neurons; Oligodendroglia; Parkinsonian Disorders; Pattern; Peripheral Nervous System; Population; Preclinical Drug Evaluation; Property; Protocols documentation; Rattus; Recovery; Regenerative Medicine; Reporter; Reporting; Research; Rodent; Science; Signal Pathway; Source; Spinal; Staging; Stem cells; Structure; Testing; Time; Transgenic Organisms; Transplantation; Tretinoin; Work; base; candidate marker; cell type; design; dopaminergic neuron; embryonic stem cell; human embryonic stem cell; human embryonic stem cell transplantation; in vivo; insight; molecular marker; nerve stem cell; neural patterning; neural plate; novel; potency testing; prospective; relating to nervous system; self-renewal; stem cell biology; stem cell population; tool; Address; Adult; Affect; Anterior; Apical; Ascorbic Acid; Astrocytes; Biological Assay; Biology; Brain; Cell Therapy; Cell physiology; Cells; Cues; Data; Derivation procedure; Development; Dopamine; Engraftment; Epiblast; Epigenetic Process; Erinaceidae; Exhibits; FGF8 gene; Fibroblast Growth Factor 2; Gene Transfer Techniques; Genes; Genetic; Glutamates; Goals; Heterogeneity; Human; In Vitro; Individual; Interneurons; Intrinsic factor; Maintenance; Midbrain structure; Molecular; Motor Neurons; Nature; Nervous system structure; Neural Crest; Neuroepithelial; Neurons; Oligodendroglia; Parkinsonian Disorders; Pattern; Peripheral Nervous System; Population; Preclinical Drug Evaluation; Property; Protocols documentation; Rattus; Recovery; Regenerative Medicine; Reporter; Reporting; Research; Rodent; Science; Signal Pathway; Source; Spinal; Staging; Stem cells; Structure; Testing; Time; Transgenic Organisms; Transplantation; Tretinoin; Work; base; candidate marker; cell type; design; dopaminergic neuron; embryonic stem cell; human embryonic stem cell; human embryonic stem cell transplantation; in vivo; insight; molecular marker; nerve stem cell; neural patterning; neural plate; novel; potency testing; prospective; relating to nervous system; self-renewal; stem cell biology; stem cell population; tool

: Neural stem cells (NSCs) can be propagated in vitro for extensive periods of time while retaining the ability to differentiate into neurons, astrocytes and oligodendrocytes. However, NSCs are limited in their potential to yield specific neuron types. Na¿ve NSCs expanded in vitro give rise primarily to GABAergic interneurons and to a lesser extent glutamate neurons. This suggests that NSCs, while multipotent, may not have access to the full spectrum of neuron types. In contrast human embryonic stem cells (hESCs) differentiated towards early neural fates can be readily biased towards various region-specific neuron types such as midbrain dopamine neurons or spinal motoneurons. We have recently reported that hESC derived neural progeny responsive to such regional patterning cues are organized into columnar neuroepithelial structures termed neural rosettes (R- NSCs) and characterized R-NSCs in considerable detail1. Our study has identified R-NSCs as novel, unique NSC stage based on marker expression, clonal stem cell properties, neural differentiation potential, and genetic identity1, 2. The most intriguing finding of these studies was the broad patterning potential of R-NSCs compared to other currently available NSC types. R-NSCs are capable of comprehensive differentiation towards CNS1 and PNS3 derivatives and capable of in vivo engraftment. Despite these exciting preliminary data our studies also revealed major gaps in our current understanding of RNSC biology. Here we would like to address some of these limitations by defining heterogeneity within RNSCs and develop genetic strategies for the prospective isolation of fully patternable R-NSCs based on BAC transgenesis. BAC transgenic hESC reporter lines have been recently pioneered in the Studer lab and will serve as reliable readout of R-NSC stage, identity and function. BAC transgenic reporters will also be critical for probing function of extrinsic and intrinsic factors affecting R-NSC identity. These studies should provide fundamental insights into the genetic and epigenetic mechanisms of neural patterning and ultimately result in novel conditions for the continued in vitro expansion of fully patternable R-NSC - a key step towards establishing a stable expandable universal NSC population.

： 神经干细胞（NSC）可以在大量时间内在体外传播，同时保持能力 分化为神经元，星形胶质细胞和少突胶质细胞。但是，NSC的潜力有限 产生特定的神经元类型。 Na¿ve nsc在体外扩展会产生gabaergic中间神经元的主要产生 较小的谷氨酸神经元。这表明NSC虽然多务件，但可能无法访问 神经元类型的光谱。相反，人类胚胎干细胞（HESC）与早期神经元区分开 命运很容易偏向各种区域特异性神经元类型，例如中脑多巴胺神经元 或脊柱运动神经元。我们最近报告说，hESC衍生出对这种反应的神经后代 区域模式提示被组织到柱神经上皮结构中，称为神经念珠（R-） NSC）并以相当大的详细说明了R-NSC。我们的研究已将R-NSC确定为新颖，独特 NSC阶段基于标记表达，克隆干细胞特性，神经分化潜力和 遗传身份1，2。这些研究最有趣的发现是R-NSC的广泛模式潜力 与其他当前可用的NSC类型相比。 R-NSC能够全面差异化 朝向CNS1和PNS3衍生物，并能够在体内植入。 尽管这些令人兴奋的初步数据我们的研究也揭示了我们当前对 RNSC生物学。在这里，我们想通过定义内在的异质性来解决其中的一些限制 RNSC并制定基于BAC的完全可模式的R-NSC的前瞻性隔离遗传策略 转带。 BAC转基因hESC报告基线最近在Studer Lab中进行了开创性，将 用作R-NSC阶段，身份和功能的可靠读数。 BAC转基因记者也将很关键 用于探测影响R-NSC身份的外在和内在因素。这些研究应提供 对神经模式的遗传和表观遗传机制的基本见解，并最终导致 持续体外扩展完全可模式的R -NSC的新型条件 - 迈向的关键步骤 建立稳定的可扩展通用NSC人群。