Characterized Adult Primary Human Microglia Cells for Research

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 与国内外研究人员合作，从正常和患病的人脑中提取通道。高产 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。大量细胞的可用性 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 RNA 和/或蛋白质的量。小胶质细胞具有多种功能 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大脑中分离的细胞。然而，较高 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 男人。研究成人大脑中小胶质细胞的功能，其中许多炎症和抗炎细胞 小胶质细胞反应的发生，需要使用成人大脑中的人类小胶质细胞。小胶质细胞培养自 人类胚胎大脑表现出巨大的增殖能力。然而，虽然隔离方法 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，从每个病例中分离和培养的小胶质细胞。我们开发了一个 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。我们建立了初级成人图书馆 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。拟议工作的具体目标是： 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，并分享来自 与国内外研究人员进行适当的交流。目标 2 – 继续表型 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 应收件人要求。目标 3 – 通过从更多案例中进行培养来补充和扩展图书馆 从现有病例中传代小胶质细胞。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。

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.