Shaqinah Nurrul Nasruddin, Mazlina Mazlan, Zamri-Saad M, Hazilawati Hamzah, Jasni Sabri
Faculty of Veterinary Medicine, Research Centre for Ruminant Diseases, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Cytotechnology. 2016 Aug;68(4):1655-9. doi: 10.1007/s10616-014-9836-7. Epub 2014 Dec 16.
Monocytes are widely used for immunological research, especially in the study of innate immune system. Although methods for isolation of human monocytes have been established, the procedure for non-human monocyte has not been well developed. This paper describes an improved method for isolation of monocyte and the subsequent macrophage cultivation from caprine blood. Monocytes were isolated from 16 ml of heparinized caprine blood using double density methods; the Ficoll and Percoll. The number of monocytes obtained was 5.12 ± 0.89 × 10(7) cells/ml at 70 % purity. The isolated monocytes were maintained in 10 % fetal bovine serum-enriched Dulbecco's Modified Eagle Medium for maturation to form macrophage cell culture. At the end of the experiment, the harvested macrophage was 2.48 ± 0.33 × 10(6) cells/ml.
单核细胞广泛应用于免疫学研究，尤其是在天然免疫系统的研究中。尽管已经建立了分离人单核细胞的方法，但非人类单核细胞的分离程序尚未得到很好的发展。本文描述了一种从山羊血液中分离单核细胞并随后培养巨噬细胞的改进方法。使用双密度法（Ficoll和Percoll）从16毫升肝素化山羊血液中分离单核细胞；获得的单核细胞数量为5.12±0.89×10⁷个细胞/毫升，纯度为70%。将分离出的单核细胞置于含有10%胎牛血清的杜氏改良 Eagle培养基中培养成熟，以形成巨噬细胞培养物。在实验结束时，收获的巨噬细胞为2.48±0.33×10⁶个细胞/毫升。
