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改进山羊模型中单核细胞的分离方法并建立巨噬细胞培养体系。

Improving the methods for isolation of monocyte and establishing macrophage cell culture in caprine model.

作者信息

Shaqinah Nurrul Nasruddin, Mazlina Mazlan, Zamri-Saad M, Hazilawati Hamzah, Jasni Sabri

机构信息

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.

出版信息

Cytotechnology. 2016 Aug;68(4):1655-9. doi: 10.1007/s10616-014-9836-7. Epub 2014 Dec 16.

Abstract

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.

摘要

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。

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Isolation of non-activated monocytes from human umbilical cord blood.从人脐血中分离非激活单核细胞。
Am J Reprod Immunol. 2010 Jan;63(1):66-72. doi: 10.1111/j.1600-0897.2009.00767.x.

本文引用的文献

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Monocyte-mediated defense against microbial pathogens.单核细胞介导的针对微生物病原体的防御。
Annu Rev Immunol. 2008;26:421-52. doi: 10.1146/annurev.immunol.26.021607.090326.
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Immunobiology of dendritic cells.树突状细胞的免疫生物学
Annu Rev Immunol. 2000;18:767-811. doi: 10.1146/annurev.immunol.18.1.767.

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