Isolation of mononuclear cells from CNS tissue using Percoll gradients centrifugation
10.3969/j.issn.1001-1978.2018.07.026
- VernacularTitle:Percoll密度梯度离心法分离中枢神经系统组织中的单个核细胞
- Author:
Cui LYU
1
;
Jin-Hong FENG
;
Zhao-Hua HOU
;
Yun-Bo WEI
;
Ting-Ting HAN
Author Information
1. 齐鲁工业大学(山东省科学院)
- Keywords:
Percoll gradients centrifugation;
mice;
central nervous system;
mononuclear cell;
flow cytometry;
lymphocyte
- From:
Chinese Pharmacological Bulletin
2018;34(7):1030-1036
- CountryChina
- Language:Chinese
-
Abstract:
Aim To establish a rapid method to efficiently iso-late mononuclear cells from central nervous system ( CNS) tis-sues of mice that can be effectively utilized for identification of various immune cell populations in a single sample by flow cy-tometry. Methods For defining the feasibility and practicality of the method, wild-type C57BL/6 mice and two mouse models of CNS disease including EAE mice and APP/PS1 mice were used in this study. After the collection and homogenization of the brain and spinal cord tissues respectively, the mononuclear cells were isolated by spinning the 70% -30% Percoll gradients. Cell activities were detected by trypan blue staining, and the im-mune population that infiltrated CNS was identified by flow cy-tometry. Results The results of trypan blue staining showed that the survival rate of the isolated cells was above 90% in all groups. Flow cytometry analysis showed that the relative num- bers of lymphocytes infiltrating CNS of EAE and AD mice in-creased significantly compared with wild-type C57BL/6 mice. In addition, the relative numbers of Th1 and Th17 cell subsets me-diating the inflammatory response also increased significantly, while the decreased regulatory T cells frequency was observed in the two mouse models of CNS disease. Conclusions The cells isolated by the 70% ~30% Percoll gradients centrifugation can be effectively utilized for the identification of various immune cell populations in a single sample by flow cytometry. The meth-od described in this article is simple and rapid in operation and with high survival rate and activity of the cells, which can be ap-plied to the study of the mononuclear cells in CNS.
