Simic Damir, Euler Catherine, Thurby Christina, Peden Mike, Tannehill-Gregg Sarah, Bunch Todd, Sanderson Thomas, Van Vleet Terry
Drug Safety Evaluation, Bristol-Myers Squibb Co, Mount Vernon, Indiana, USA.
Curr Protoc Toxicol. 2012 Aug;Chapter 14:Unit 14.9.1-17. doi: 10.1002/0471140856.tx1409s53.
In this in vitro model of hepatocyte multinucleation, separate cultures of rat Clone 9 cells are labeled with either red or green cell tracker dyes (Red Cell Tracker CMPTX or Vybrant CFDA SE Cell Tracer), plated together in mixed-color colonies, and treated with positive or negative control agents for 4 days. The fluorescent dyes become cell-impermeant after entering cells and are not transferred to adjacent cells in a population, but are inherited by daughter cells after fusion. The mixed-color cultures are then evaluated microscopically for multinucleation and analysis of the underlying mechanism (cell fusion/cytokinesis). Multinucleated cells containing only one dye have undergone cytokinesis failure, whereas dual-labeled multinucleated cells have resulted from fusion.
在这个肝细胞多核化的体外模型中，将大鼠克隆9细胞的单独培养物用红色或绿色细胞追踪染料（红细胞追踪剂CMPTX或Vybrant CFDA SE细胞示踪剂）进行标记，一起接种形成混合颜色的集落，并用阳性或阴性对照剂处理4天。荧光染料进入细胞后变得不能透过细胞膜，并且不会转移到群体中的相邻细胞，但在融合后会被子代细胞继承。然后通过显微镜对混合颜色培养物进行多核化评估，并分析其潜在机制（细胞融合/胞质分裂）。仅含有一种染料的多核细胞经历了胞质分裂失败，而双标记的多核细胞则是融合的结果。
