An optimized method for the induction and purification of mouse bone marrow dendritic cells

Liu, Ling; Fan, Shanwen; Lu, Zhonghua; Chen, Zhenxing; Chu, Cuilin; Liu, Airan; Xia, Feiping; Meng, Shanshan; Guo, Fengmei; Qiu, Haibo; Yang, Yi

doi:10.1016/j.jim.2021.113073

Cited by 3 publications

(1 citation statement)

References 31 publications

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“…These debris make the isolation of cells from muscle less efficient and were therefore removed using Percoll density gradient centrifugation [ 35 , 36 ] and fluorescence-activated cell sorting [ 37 ]. The Percoll gradient method is widely used for isolating lymphocytes from tissues other than the muscles [ 38 , 39 , 40 ]. The CD45-magnetic bead isolation method is another alternative for lymphocyte isolation [ 41 , 42 , 43 ].…”

Section: Introductionmentioning

confidence: 99%

Efficient Isolation of Lymphocytes and Myogenic Cells from the Tissue of Muscle Regeneration

Kitajima

Tsukahara

Nakamoto

et al. 2022

Cells

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Isolation of both lymphocytes and myogenic cells from muscle tissue is required for elucidating the cellular and molecular mechanisms of muscle regeneration. Here, we aimed to establish an optimal method obtaining a high yield of lymphocytes during muscle regeneration. After the muscle injury, we observed higher infiltration of lymphocytic cells in the muscle on day 3 after injury. Then, we compared two different white blood cell isolation methods, the Percoll gradient and CD45-magnetic bead methods, to assess the percentage and number of T and B cells. Flow cytometry analysis showed that the CD45-magnetic bead method has a better efficiency in isolating CD4+, CD8+ T cells, and B cells from injured muscle tissues of wild-type and mdx mice than that by the Percoll gradient method. Moreover, we found that the CD45-negative fraction from wild-type and mdx mice includes myogenic cells. In conclusion, we report that the CD45-magnetic bead method is suitable to isolate T and B cells during muscle regeneration with higher purity and yield and can also isolate myogenic cells within the same sample. This method provides a technical basis for further studies on muscle regeneration, involving lymphocytes and muscle cells, with a wide range of clinical applications.

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