Abstract:
Tissues are formed and shaped by cells of many different types and are orchestrated through countless interactions. Deciphering a tissue\'s biological complexity thus requires studying it at cell-level resolution, where molecular and biochemical features of different cell types can be explored and thoroughly dissected. Unfortunately, the lack of comprehensive methods to identify, isolate, and culture each cell type from many tissues has impeded progress. Here, we present a method for the breadth of cell types composing the human breast. Our goal has long been to understand the essence of each of these different breast cell types, to reveal the underlying biology explaining their intrinsic features, the consequences of interactions, and their contributions to the tissue. This biological exploration has required cell purification, deep-RNA sequencing-and a thorough dissection of the genes and pathways defining each cell type. Whereas the molecular analysis is presented in an adjoining article, we present here an exhaustive cellular dissection of the human breast and explore its cellular composition and histological organization. Moreover, we introduce a novel FACS antibody panel and rigorous gating strategy capable of isolating each of the twelve major breast cell types to purity. Finally, we describe the creation of primary cell models from nearly every breast cell type-some the first of their kind- and submit these as critical tools for studying the dynamic cellular interactions within breast tissues and tumors. Together, this body of work delivers a unique perspective of the breast, revealing insights into its cellular, molecular, and biochemical composition.
摘要:
组织由许多不同类型的细胞形成和塑造,并通过无数的相互作用进行编排。因此,破译组织的生物复杂性需要以细胞水平的分辨率来研究它,可以探索和彻底解剖不同细胞类型的分子和生化特征。不幸的是,缺乏全面的识别方法,隔离,从许多组织中培养每种细胞类型阻碍了进展。这里,我们提出了一种用于构成人类乳房的细胞类型的广度的方法。我们的目标一直是了解这些不同乳腺细胞类型的本质,来揭示解释其内在特征的潜在生物学,相互作用的后果,以及它们对组织的贡献。这种生物探索需要细胞纯化,深度RNA测序-以及定义每种细胞类型的基因和途径的彻底解剖。而分子分析在相邻的文章中提出,我们在这里介绍了人类乳房的详尽细胞解剖,并探讨了其细胞组成和组织学组织。此外,我们引入了一种新的FACS抗体组和严格的门控策略,能够分离12种主要乳腺细胞类型中的每一种至纯度.最后,我们描述了从几乎所有乳腺细胞类型中创建原代细胞模型的过程-有些是它们的第一个类型-并将这些模型作为研究乳腺组织和肿瘤内动态细胞相互作用的关键工具。一起,这项工作提供了独特的乳房视角,揭示了对其细胞的洞察力,分子,和生化成分。