Characterized Adult Primary Human Microglia Cells for Research

拟议项目的目标是分享各种成人原代人类小胶质细胞的特征， 与国内外的研究人员一起从正常和患病的人类大脑中提取基因片段。高产 并且我们的文库中的成熟的人类小胶质细胞培养物将允许体外研究的机制 在不同的实验条件下使用来自同一病例的细胞。大量电池的可用性 还将允许对可以调节小胶质细胞功能的药物进行初步测试，并允许分离大胶质细胞。 量的RNA和/或蛋白质用于转录组学或蛋白质组学研究。小胶质细胞在脑内具有多种功能， 健康和患病的大脑关于小胶质细胞生物学的大量知识都是基于 体外研究绝大多数使用从啮齿动物脑中分离的细胞。然而， 人类大脑的解剖和功能复杂性以及小胶质细胞反应的物种差异， 功能使迫切需要使用人类小胶质细胞，以确定所获得的结果适用于 伙计调查小胶质细胞在成人大脑中的功能，其中许多炎症和抗炎 小胶质细胞反应的发生，需要使用来自成人大脑的人类小胶质细胞。培养的小胶质细胞 胚胎人脑显示出相当大的增殖能力。然而，虽然用于分离的方法 虽然已经描述了成年人死后大脑中的小胶质细胞，但他们只允许使用有限的数量 由于增殖水平低，从每个病例中分离和培养的小胶质细胞的数量。我们已经开发出一种 一种允许从成年死后人脑中培养小胶质细胞到高传代的方法， 这些细胞在高传代培养物中保持其表型。我们建立了一个初级成人图书馆， 来自年轻和老年死后脑以及来自 患有各种神经退行性疾病的个体。这项工作的主要目标是分享 人类小胶质细胞的研究。拟议工作的具体目标是： 关于从正常年轻人脑中获得表征的成人原代人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， 适当的通道与研究人员在国内和国际。目标2 -延续表型 人小胶质细胞培养物的表征，包括深度测序，以及作为 收件人要求。目标3 -通过从额外的病例培养来补充和扩大文库， 从现有病例中传代小胶质细胞。分享来自不同病例的初级成人小胶质细胞， 不同的通道将为科学界提供一个新的和可靠的工具，体外机制， 研究人类小胶质细胞在从儿童到老年的健康中的功能，以及在神经系统疾病中的功能。

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.