Characterized Adult Primary Human Microglia Cells for Research

该项目的目标是共享不同类型的成人原代小胶质细胞。 国内外研究人员从正常和患病的人脑中摘录的文章。高产 来自我们图书馆的具有良好特性的成人小胶质细胞培养将允许机械性的体外研究 同一病例的细胞在不同的实验条件下使用。大量单元格的可用性 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，并允许分离出 用于转录组或蛋白质组学研究的RNA和/或蛋白质的量。小胶质细胞在血管内皮细胞中具有多种功能。 健康和患病的大脑。许多关于小胶质细胞生物学的知识都是基于 体外研究中的绝大多数使用了从啮齿动物大脑中分离出来的细胞。然而，更高的 人脑解剖和功能的复杂性以及小胶质细胞反应和 功能使得使用人类小胶质细胞来确定所获得的结果适用于 天哪。成人脑内多种炎症和抗炎状态下小胶质细胞功能的研究 小胶质细胞的反应需要使用成人大脑中的人小胶质细胞。培养的小胶质细胞 胚胎人脑显示出较强的增殖能力。然而，虽然分离的方法 已经描述了成人死后大脑中的小胶质细胞，它们只允许有限数量的使用 由于低水平的增殖，从每个病例分离和培养的小胶质细胞。我们已经开发出一种 一种允许将成人死后脑小胶质细胞培养到高传代的方法，并发现了 这样的细胞在高传代培养中保持其表型。我们已经建立了一个初级成人图书馆 人不同代次的小胶质细胞，分别取自青年和老年死后脑和 患有各种神经退行性疾病的人。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 关于从正常年轻人的大脑中获得具有特征的成人原代人类小胶质细胞的信息， 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 与国内和国际研究人员进行适当交流。目标2--持续表型 人类小胶质细胞培养的特征，包括深度测序，以及其他特征，如 由收件人请求。目标3-通过培养更多病例和 从现有病例中传代小胶质细胞。不同情况下原代成人小胶质细胞的共享性 不同的通道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儿童到老年的健康和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.