A nano-enabled biomimetic platform for neuronal differentiation and maturation

用于神经元分化和成熟的纳米仿生平台

基本信息

批准号：
9809234
负责人：
Larry J. Millet
金额：
$ 7.55万
依托单位：
UNIVERSITY OF TENNESSEE KNOXVILLE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9809234
关键词：
Alcoholism Ammonia Animals Axon Biochemical Biological Assay Biology Biomimetics Brain Brain region Cell Culture Techniques Cells Chemicals Coculture Techniques Complex Cues Data Dense Core Vesicle Development Diabetes Mellitus Disease Disease model Drug Screening Exhibits Exocytosis Extracellular Matrix Foundations Future Glass Goals Growth Growth and Development function Hormones Human Image Image Analysis Improve Access In Vitro Instruction Label Lead Ligands Maps Measurement Measures Mental Depression Metabolic Methods Microscopy Millet Modeling Modernization Molecular Nanotechnology Nanotopography Neuroglia Neuronal Differentiation Neurons Neurosciences Neurosecretory Systems Neurotransmitters Oxytocin Pattern Pharmaceutical Preparations Pharmacogenomics Pharmacotherapy Polylysine Population Process Production Psychiatric therapeutic procedure Reagent Regulation Resources Risk Management Signal Transduction Silicon Spectrum Analysis Stem cells Surface Technology Testing Time Tissue Microarray Tissues Transfection Translational Research Vasopressins Work autism spectrum disorder cell type cost data acquisition density drug candidate drug discovery drug testing effectiveness measure experience experimental study extracellular high resolution imaging imaging study improved in vivo induced pluripotent stem cell innovation interstitial magnocellular nano nanoscale neurochemistry novel post-traumatic stress prevent receptor silicon nitride skills translational medicine validation studies

项目摘要

Project Summary Access to mature cells and tissues is key to understanding the pathobiology of diseases and for developing drug therapies. Cellular and molecular studies of nerve cells are a cornerstone of modern neuroscience, yet magnocellular neurons (MCN) and some stem cell types defy conventional culture methods. For over 20 years, the challenge of culturing MCNs in vitro has been an obstacle for cellular and molecular studies on the production and regulation of oxytocin and vasopressin hormones, neurotransmitters, and their receptors. For unknown reasons, MCN cannot be grown in vitro without feeder layers. Conventional substrates have not yet been able to replace these co-dependent requirements. The goal of this proposal is to harness physical contacts and chemical signatures of the extracellular matrix to create a cell culture platform that reliably promotes, maintains, and matures MCN in vitro. We describe a two-stage approach that maximizes the progress and manages the risks while advancing innovation to solve the MCN culture challenge. First, we will test and compare brain decellularized extracellular matrix (Brain-DECM) to conventional methods to improve MCN yields beyond the established co-culture model. This will substantially reduce the time, variability, and complexity of experiments by refining the cell culture process to replace feeder-layers and decrease animal use. Because Brain-DECM is complex reagent with many unknowns, our second stage is to create a nano-scale biomimetic glial-derived extracellular matrix. The result is a tissue chip that retains the growth-permissive physical cues while incorporating developmentally-instructive biochemical ligands derived from glia. This tissue chip platform may improve neuron yield and increase access to mature MCNs to enable high-resolution imaging and analyses for difficult to culture cellular populations. We expect that this innovative solution to a long- standing challenge will be reliable, productive, and impactful to yield new opportunities for drug discovery for translational science and medicine.

项目摘要进入成熟的细胞和组织是理解疾病病理生物学和发展的关键药物疗法。神经细胞的细胞和分子研究是现代神经科学的基石，但巨细胞神经元（MCN）和一些干细胞类型反对常规培养方法。超过20年，体外培养MCN的挑战一直是细胞和分子研究的障碍催产素和加压素激素，神经递质及其受体的生产和调节。为了未知的原因，没有进料器层，MCN不能在体外生长。传统的基材尚未能够替代这些共同依赖的要求。该提议的目的是利用身体接触以及细胞外基质的化学特征，以创建一个可靠地促进的细胞培养平台维持并在体外成熟MCN。我们描述了一种两阶段的方法，可最大化进度和管理风险，同时推进创新以解决MCN文化挑战。首先，我们将测试和比较脑脱细胞外基质（脑decm）与改善MCN的常规方法产量超出了既定的共培养模型。这将大大减少时间，可变性和通过完善细胞培养过程来替代进料器并减少动物的使用，实验的复杂性。因为脑decm是复杂的试剂，但我们的第二阶段是创建纳米级仿生神经胶质细胞外基质。结果是保留生长腐蚀性的组织芯片物理线索同时结合了源自胶质的发展的结构生化配体。这个组织芯片平台可以提高神经元的产量并增加获得成熟MCN的访问，以实现高分辨率成像并分析难以培养细胞种群。我们期望这种创新的解决方案可以长期站立挑战将是可靠，富有成效和影响力的，以产生新的毒品发现机会翻译科学与医学。