Cell Cycle Analysis by Mass Cytometry
Cell Cycle Analysis by Mass Cytometry
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DOI：
10.1007/978-1-4939-7371-2_8
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Behbehani, Gregory K.
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文献类型：
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作者：
Behbehani, Gregory K.
细胞在增殖期间通过细胞周期的调节进展是肿瘤进展、抗肿瘤治疗应答、免疫系统调节和发育生物学中的关键因素。虽然细胞周期进程的流式细胞术测量是完善的，但质谱细胞术测定允许细胞周期与多达39种其他抗原沿着测量，从而能够表征细胞周期与各种细胞过程之间的复杂相互作用。该方法描述了使用质谱细胞术分析来自三种不同来源的细胞的细胞周期状态：体外培养的细胞系、离体人血液或骨髓以及小鼠组织的体内标记和离体分析。该方法利用5-碘-2 '-脱氧尿苷（IdU）掺入，并结合测量磷酸化视网膜母细胞瘤蛋白（pRb）、细胞周期蛋白B1和磷酸化组蛋白H3（p-HH 3）。这些测量可以集成到门控策略中，该策略允许清楚地分离细胞周期的所有五个阶段。
The regulated progression of cells through the cell cycle during proliferation is a critical factor in tumor progression, anti-neoplastic therapy response, immune system regulation, and developmental biology. While flow cytometric measurement of cell cycle progression is well established, mass cytometry assays allow the cell cycle to be measured along with up to 39 other antigens enabling characterization of the complex interactions between the cell cycle and a wide variety of cellular processes. This method describes the use of mass cytometry for the analysis of cell cycle state for cells from three different sources: in vitro cultured cell lines, ex vivo human blood or bone marrow, and in vivo labeling and ex vivo analysis of murine tissues. The method utilizes incorporation of 5-Iodo-2'-deoxyuridine (IdU), combined with measurement of phosphorylated retinoblastoma protein (pRb), cyclin B1, and phosphorylated histone H3 (p-HH3). These measurements can be integrated into a gating strategy that allows for clear separation of all five phases of the cell cycle.