Laboratory of Parasitic Diseases, National Institutes of Health, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA
Infect Immun. 2019 Oct 18;87(11). doi: 10.1128/IAI.00639-19. Print 2019 Nov.
is usually cultured axenically in TYI-S-33, a complex medium which does not permit survival and growth of mammalian cells. Likewise, medium commonly used to maintain and grow mammalian cells does not support healthy trophozoite survival for more than a few hours. The inability to coculture trophozoites and epithelial cells under optimal conditions limits studies of their interactions as well as interpretation of results. Trophozoites of the WB isolate but not the GS isolate were repeatedly adapted to grow stably in long-term cocultures with Caco2, Cos7, and mouse tumor rectal (RIT) cell lines using hybridoma-screened Dulbecco's modified Eagle's medium and 10% fetal calf serum. did not grow in spent cell culture medium or when separated by a permeable membrane using transwell methodology. chronically cocultured with specific cell lines became adapted (conditioned). These cocultures grew better than nonconditioned trophozoites, and the cell lines differed in their ability to support trophozoite growth in the order of RIT > Cos7 > Caco2. Trophozoites conditioned on one cell line and then grown in the presence of a heterologous cell line changed their growth rate to that seen in conditioned from the heterologous cell line. Trophozoite survival required intimate contact with cells, suggesting that trophozoites obtain an essential nutrient or growth factor from mammalian cells. This may explain why trophozoites adhere to the small intestinal epithelium during human and animal infections. This coculture system will be useful to understand the complex interactions between the host cells and parasite.
通常在 TYI-S-33 中进行无细胞培养,这是一种复杂的培养基,不允许哺乳动物细胞存活和生长。同样,用于维持和生长哺乳动物细胞的培养基也不能支持滋养体在数小时以上的健康存活。由于无法在最佳条件下共培养滋养体和上皮细胞,限制了对它们相互作用的研究以及对结果的解释。WB 分离株的滋养体但不是 GS 分离株的滋养体被反复适应在长期与 Caco2、Cos7 和鼠直肠 (RIT) 细胞系的共培养中稳定生长,使用杂交瘤筛选的 Dulbecco 改良 Eagle 培养基和 10%胎牛血清。滋养体不会在耗尽的细胞培养物中生长,也不会在使用 Transwell 方法通过可渗透膜分离时生长。与特定细胞系慢性共培养的滋养体适应(条件化)。这些共培养物比非条件化的滋养体生长得更好,并且细胞系在支持滋养体生长的能力方面存在差异,顺序为 RIT > Cos7 > Caco2。在存在异源细胞系的情况下,在一种细胞系上条件化的滋养体改变其生长速度,达到来自异源细胞系的条件化滋养体的生长速度。滋养体的存活需要与细胞的密切接触,这表明滋养体从哺乳动物细胞中获得必需的营养或生长因子。这可以解释为什么在人类和动物感染期间,滋养体附着在小肠上皮上。这种共培养系统将有助于理解宿主细胞和寄生虫之间的复杂相互作用。