Wang Ruoxi W, MacDuffie Emily, Santaguida Stefano
Koch Institute for Integrative Cancer Research at MIT, Department of Biology, Massachusetts Institute of Technology.
Koch Institute for Integrative Cancer Research at MIT, Department of Biology, Massachusetts Institute of Technology;
J Vis Exp. 2018 Apr 13(134):57215. doi: 10.3791/57215.
Chromosome mis-segregation leads to aneuploidy, a condition in which cells harbor an imbalanced chromosome number. Several lines of evidence strongly indicate that aneuploidy triggers genome instability, ultimately generating cells with complex karyotypes that arrest their proliferation. Isolation and characterization of cells harboring complex karyotypes are crucial to study the impact of an imbalanced chromosome number on cell physiology. To date, no methods have been established to reliably isolate such aneuploid cells. This paper provides a protocol for the enrichment and analysis of aneuploid cells with complex karyotypes utilizing standard, inexpensive tissue culture techniques. This protocol can be used to analyze several features of aneuploid cells with complex karyotypes including their induced senescence-associated secretory phenotype, pro-inflammatory properties, and ability to interact with immune cells. Because cancer cells often harbor imbalances in chromosome number, it is crucial to decipher how aneuploidy impacts cell physiology in normal cells, with the ultimate goal of uncovering both its pro- and anti-tumorigenic effects.
染色体错分离会导致非整倍体，即细胞中染色体数目失衡的一种状态。多条证据有力地表明，非整倍体引发基因组不稳定，最终产生具有复杂核型的细胞，这些细胞会停止增殖。分离和鉴定具有复杂核型的细胞对于研究染色体数目失衡对细胞生理学的影响至关重要。迄今为止，尚未建立可靠地分离此类非整倍体细胞的方法。本文提供了一种利用标准、廉价的组织培养技术富集和分析具有复杂核型的非整倍体细胞的方案。该方案可用于分析具有复杂核型的非整倍体细胞的多个特征，包括其诱导的衰老相关分泌表型、促炎特性以及与免疫细胞相互作用的能力。由于癌细胞常常存在染色体数目失衡，因此解读非整倍体如何影响正常细胞的细胞生理学至关重要，最终目标是揭示其促肿瘤和抗肿瘤作用。
