Drastic changes in chromosome number and cellular contents upon ploidy alterations profoundly affect the stability of mitotic regulation in different biological and pathological processes. Isogenic ploidy series of somatic cell lines are useful for studying the effects of ploidy differences on mitotic regulation at cellular and molecular levels. This chapter describes experimental procedures using isogenic human HAP1 cell lines that cover haploid, diploid, and tetraploid states. We first describe methods to establish and maintain these isogenic HAP1 ploidy series using a flow cytometer. We then describe a procedure of comparative pharmacological assay for analyzing ploidy-dependent changes in the functionality of the mitotic spindle components.
Keywords: Centrosome; Chromosome segregation; Flow cytometry; HAP1 cells; Mitotic spindle; Ploidy; Size scaling; Whole-genome duplication.
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