Abstract：
Polyploid giant cancer cells (PGCCs) play a fundamental role in tumor initiation, dormancy, drug resistance, and metastasis, although the detailed biology of PGCCs remains poorly understood. The lack of literature on establishing a reproducible in vitro system for generating PGCCs is the leading technological obstacle to studying the biology of PGCCs. Here we provide a detailed protocol for generating stable PGCCs from Hey cancer cells and studying the PGCCs\' embryonic stemness. This protocol includes (1) generating PGCCs of high purity in 2D culture by exposing Hey cells to paclitaxel, monitoring the cell cycle and amitotic budding of daughter cells from PGCCs, and collecting and studying the daughter cells; (2) inducing PGCCs to form spheroids expressing embryonic stemness markers and observing the spheroids\' cleavage and blastocyst-like structure; and (3) inducing redifferentiation of PGCCs into different lineages of differentiated cells.
摘要：
多倍体巨癌细胞(PGCCs)在肿瘤的发生、发展中起着重要作用,休眠,耐药性,和转移，尽管PGCC的详细生物学仍知之甚少。缺乏关于建立用于产生PGCC的可重复体外系统的文献是研究PGCC生物学的主要技术障碍。在这里，我们提供了从Hey癌细胞产生稳定的PGCCs和研究PGCCs胚胎干细胞的详细方案。该方案包括（1）通过将Hey细胞暴露于紫杉醇在2D培养物中产生高纯度的PGCC，监测PGCC子细胞的细胞周期和有丝分裂出芽，并收集和研究子细胞；（2）诱导PGCC形成表达胚胎干细胞标记的球体，并观察球体的卵裂和囊胚样结构；（3）诱导PGCC再分化为不同谱系的分化细胞。
