Characterized Adult Primary Human Microglia Cells for Research

用于研究的特征化成人原代人小胶质细胞

基本信息

批准号：
9788539
负责人：
CHANGIZ GEULA
金额：
$ 26.3万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9788539
关键词：
Adult Age Anatomy Anti-inflammatory Autopsy Biology Brain Brain Diseases Cells Cerebral cortex Childhood Communities Disease Drug Screening Embryo Ensure Explosion Extracellular Matrix Free Radicals Goals Health Human Immune In Vitro Individual Inflammatory Injections International Investigation Libraries Maintenance Methods Microglia Myelogenous Neuraxis Neurodegenerative Disorders Neurons Nitric Oxide Pathology Phenotype Physiological Primates Process Proteins Proteomics RNA Rattus Research Research Personnel Resource Sharing Resources Rodent Rodent Model Role Stimulus Synapses Testing Trauma Traumatic Brain Injury Work aged amyloid peptide base brain cell brain tissue deep sequencing drug testing human disease in vivo interest macrophage man nervous system development nervous system disorder neuropathology nonhuman primate response species difference tool transcriptomics

项目摘要

The goal of the proposed project is to share characterized adult primary human microglia cells of various passages from normal and diseased human brains with researchers nationally and internationally. High yield and well characterized adult human microglia cultures from our library will allow mechanistic in vitro studies with cells from the same case used in different experimental conditions. Availability of large numbers of cells would also allow initial testing of drugs that can regulate microglial functions and permit isolation of large amounts of RNA and/or protein for transcriptomic or proteomic studies. Microglia have diverse functions in the healthy and diseased brain. A great deal of what has been learned regarding microglia biology is based on in vitro studies the overwhelming majority of which used cells isolated from the rodent brain. However, higher anatomical and functional complexity of the human brain and species differences in microglia response and function make imperative the use of human microglia to ascertain that the results obtained are applicable to man. Investigation of microglia function in the adult brain, in which many inflammatory and anti-inflammatory microglia responses occur, requires use of human microglia from adult brains. Microglia cultured from embryonic human brain show substantial proliferative capacity. However, while methods for isolation of microglia from adult postmortem human brains had been described, they allowed only use of a limited quantity of microglia isolated and cultured from each case due to low levels of proliferation. We have developed a method that allows culturing microglia from adult postmortem human brains to high passage and have found that such cells maintain their phenotype in high passage cultures. We have built a library of primary adult human microglia cells of various passages from young and aged postmortem brains and from brains of individuals with various neurodegenerative disorders. The primary goal of the proposed work is to share human microglia with researchers. The specific aims of the proposed work are: Aim 1 – Disseminate information regarding availability of characterized adult primary human microglia from brains of normal young, normal aged and brains with various neurodegenerative disorders for research and share such microglia from appropriate passages with researchers nationally and internationally. Aim 2 – Continue phenotypic characterization of human microglia cultures, including deep sequencing, and additional characterization as requested by recipients. Aim 3 – Replenish and expand the library by culturing from additional cases and passaging microglia from existing cases. Sharing of primary adult human microglia from various cases and different passages will provide the scientific community with a new and reliable tool for in vitro mechanistic studies of human microglia function in health from childhood to old age, and in diseases of the nervous system.

拟议项目的目标是共享各种特征的成人原代人类小胶质细胞来自国内和国际研究人员的正常和患病人类大脑的通道高产量。我们的图书馆中经过充分表征的成人小胶质细胞培养物将允许进行机械体外研究来自同一病例的细胞在不同的实验条件下使用。还可以对能够调节小胶质细胞功能并允许分离大胶质细胞的药物进行初步测试。用于转录组或蛋白质组研究的大量 RNA 和/或蛋白质在细胞中具有多种功能。关于小胶质细胞生物学的大量知识都是基于健康和患病的大脑。体外研究绝大多数使用从啮齿动物大脑中分离出来的细胞。人脑的解剖和功能复杂性以及小胶质细胞反应和的物种差异功能使得必须使用人类小胶质细胞来确定所获得的结果适用于成人大脑中小胶质细胞功能的研究，其中许多炎症和抗炎作用。小胶质细胞反应的发生需要使用从成人大脑中培养的人类小胶质细胞。人类胚胎大脑显示出巨大的增殖能力，然而，分离方法却很困难。已经描述了来自成人死后大脑的小胶质细胞，但他们只允许使用有限数量的由于增殖水平低，我们开发了一种从每个病例中分离和培养的小胶质细胞。允许从成年死后人类大脑中培养小胶质细胞进行高传代的方法，并发现这些细胞在高传代培养物中保持其表型我们已经建立了一个原代成体文库。来自年轻人和老年人死后大脑以及来自不同阶段的人类小胶质细胞患有各种神经退行性疾病的个体拟议工作的主要目标是分享。人类小胶质细胞与研究人员合作的具体目标是：目标 1 – 传播有关来自正常年轻人大脑的特征性成人原代人类小胶质细胞的可用性的信息，正常老年人和患有各种神经退行性疾病的大脑进行研究，并共享此类小胶质细胞目标 2 – 继续研究表型。人类小胶质细胞培养物的表征，包括深度测序，以及其他表征目标 3 – 通过培养更多病例来补充和扩展库。从现有病例中传递小胶质细胞分享来自不同病例的原代成人小胶质细胞不同的段落将为科学界提供一种新的、可靠的体外机制工具研究人类小胶质细胞在从童年到老年的健康以及神经系统疾病中的功能。