Hoffman Robert M, Chishima Takashi
AntiCancer Inc., San Diego, CA, USA.
Department of Surgery, UCSD, San Diego, CA, USA.
Methods Mol Biol. 2018;1760:215-220. doi: 10.1007/978-1-4939-7745-1_20.
Distant organ colonization by cancer cells is the governing step of metastasis. We review in this chapter the modeling and imaging of organ colonization by cancer cells in Gelfoam histoculture. ANIP 973 lung cancer cells expressing green fluorescent protein (GFP) were injected intravenously into nude mice, whereby they formed brilliantly fluorescing metastatic colonies on the mouse lung. The seeded lung tissue was then excised and incubated in the three-dimensional Gelfoam histoculture that maintained the critical features of progressive in vivo organ colonization. Tumor progression was continuously visualized by GFP fluorescence of individual cultures over a 52-day period, during which tumor colonies spread throughout the lung. Organ colonization was selective in Gelfoam histoculture for lung cancer cells to grow on lung tissue, since no growth occurred on histocultured mouse liver tissue. The ability to support selective organ colonization in Gelfoam histoculture and visualize tumor progression by GFP fluorescence allows the in vitro study of the governing processes of metastasis.
癌细胞向远处器官的定植是转移的关键步骤。在本章中,我们将综述在明胶海绵组织培养中癌细胞向器官定植的建模与成像。将表达绿色荧光蛋白(GFP)的ANIP 973肺癌细胞静脉注射到裸鼠体内,这些细胞在小鼠肺部形成了发出明亮荧光的转移瘤菌落。然后切除接种了细胞的肺组织,并在三维明胶海绵组织培养中进行培养,该培养体系保留了体内器官进行性定植的关键特征。在52天的时间里,通过对单个培养物的GFP荧光持续观察肿瘤进展情况,在此期间肿瘤菌落在整个肺部扩散。在明胶海绵组织培养中,器官定植具有选择性,肺癌细胞能够在肺组织上生长,而在组织培养的小鼠肝脏组织上则不会生长。明胶海绵组织培养支持选择性器官定植并通过GFP荧光观察肿瘤进展的能力,使得对转移的关键过程进行体外研究成为可能。