Ali A, Reynolds D L
Department of Microbiology, Immunology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
Avian Dis. 1996 Jan-Mar;40(1):103-8.
Primary cell culture has been widely used in various types of studies and proven useful for the isolation and identification of avian pathogens. Difficulties in growing intestinal epithelial cells in vitro have limited their use for such studies. In the present study, a co-culture system was developed for the primary culture of intestinal epithelial cells. A monolayer obtained from 14-to- 16-day-old turkey embryo intestinal fibroblasts was used as a feeder layer. Feeder layers from turkey embryo fibroblasts and from a continuous cell line (mouse 3T3 fibroblasts) were also employed but were not as successful. The intestinal epithelial cells were isolated by dissociation from the intestinal tracts of 1-day-old turkey poults and grown on the feeder layers. Growth and maintenance media were supplemented with various components, including fetal calf serum, chicken serum, hormones, and other growth factors. The epithelial cells grown on feeder layers from the intestinal fibroblasts allowed the intestinal epithelial cells to be maintained in vitro for periods of 7 to 10 days. This technique may prove useful for various applications, including isolation of enteropathogens, and for basic studies of the intestinal tract concerning such subjects as physiology, immunology, and toxicology.
原代细胞培养已广泛应用于各类研究，并已证明其在禽病原体分离和鉴定方面很有用。体外培养肠道上皮细胞存在的困难限制了其在此类研究中的应用。在本研究中，开发了一种用于肠道上皮细胞原代培养的共培养系统。从14至16日龄火鸡胚胎肠道成纤维细胞获得的单层细胞用作饲养层。也使用了来自火鸡胚胎成纤维细胞和连续细胞系（小鼠3T3成纤维细胞）的饲养层，但效果不佳。通过从1日龄小火鸡的肠道中解离来分离肠道上皮细胞，并在饲养层上生长。生长和维持培养基补充了各种成分，包括胎牛血清、鸡血清、激素和其他生长因子。在来自肠道成纤维细胞的饲养层上生长的上皮细胞可使肠道上皮细胞在体外维持7至10天。该技术可能对包括肠道病原体分离在内的各种应用以及对肠道生理学、免疫学和毒理学等主题的基础研究有用。
