Centro de Investigacao Translacional em Oncologia (LIM24), Instituto do Cancer do Estado de 11Sao Paulo (ICESP), Sao Paulo, Brazil.
Department of Microbiology, Institute of Biomedical Sciences, University of Sao Paulo (USP), Av. Prof. Lineu Prestes 1374, 05508-900, Sao Paulo, SP, Brazil.
Curr Top Med Chem. 2018;18(4):246-255. doi: 10.2174/1568026618666180410125850.
In vitro culture of primary or established cell lines is one of the leading techniques in many areas of basic biological research. The use of pure or highly enriched cultures of specific cell types obtained from different tissues and genetics backgrounds has greatly contributed to our current understanding of normal and pathological cellular processes. Cells in culture are easily propagated generating an almost endless source of material for experimentation. Besides, they can be manipulated to achieve gene silencing, gene overexpression and genome editing turning possible the dissection of specific gene functions and signaling pathways. However, monolayer and suspension cultures of cells do not reproduce the cell type diversity, cell-cell contacts, cell-matrix interactions and differentiation pathways typical of the three-dimensional environment of tissues and organs from where they were originated. Therefore, different experimental animal models have been developed and applied to address these and other complex issues in vivo. However, these systems are costly and time consuming. Most importantly the use of animals in scientific research poses moral and ethical concerns facing a steadily increasing opposition from different sectors of the society. Therefore, there is an urgent need for the development of alternative in vitro experimental models that accurately reproduce the events observed in vivo to reduce the use of animals. Organotypic cultures combine the flexibility of traditional culture systems with the possibility of culturing different cell types in a 3D environment that reproduces both the structure and the physiology of the parental organ. Here we present a summarized description of the use of epithelial organotypic for the study of skin physiology, human papillomavirus biology and associated tumorigenesis.
原代或已建立的细胞系的体外培养是许多基础生物学研究领域的主要技术之一。使用从不同组织和遗传背景中获得的纯或高度富集的特定细胞类型培养物,极大地促进了我们对正常和病理细胞过程的当前理解。培养中的细胞易于繁殖,产生了几乎无穷无尽的实验材料来源。此外,它们可以被操纵以实现基因沉默、基因过表达和基因组编辑,从而有可能剖析特定基因功能和信号通路。然而,单层和悬浮细胞培养不能再现细胞类型多样性、细胞-细胞接触、细胞-基质相互作用和分化途径,这些是它们起源的组织和器官的三维环境的典型特征。因此,已经开发并应用了不同的实验动物模型来解决这些和其他体内的复杂问题。然而,这些系统成本高且耗时。最重要的是,在科学研究中使用动物引起了道德和伦理方面的关注,这导致社会各界的反对声音日益增加。因此,迫切需要开发能够准确再现体内观察到的事件的替代体外实验模型,以减少动物的使用。器官型培养将传统培养系统的灵活性与在 3D 环境中培养不同细胞类型的可能性相结合,该环境既再现了母体器官的结构又再现了其生理学。在这里,我们对上皮器官型培养在皮肤生理学、人乳头瘤病毒生物学和相关肿瘤发生学研究中的应用进行了简要描述。