A method for high purity sorting of rare cell subsets applied to T-DC

作者：Kuka Mirela^*; Ashwell Jonathan D

来源：Journal of Immunological Methods, 2013, 400: 111-116.

DOI：10.1016/j.jim.2013.10.002

• Flow cytometry
• Sorting
• T-DC
• Purification

T-DC are a recently described subset of polyclonal alpha beta T-cells with dendritic cell properties. Because of their low number in peripheral immune compartments, isolation and characterization of T-DC with existing purification methods are technically challenging. Here we describe a customized gating strategy and a flow cytometry-based cell sorting protocol for isolation of T-DC. The protocol was developed because, despite very conservative gating for dead-cell and doublet exclusion, cells obtained with normal sorting procedures were enriched for T-DC but not pure. Re-sorting the output of the first round of sorting results in highly pure T-DC. Cells obtained with this method are viable and can be used for in vitro characterization. Moreover, this double-round sorting strategy can be universally applied to the isolation of other rare cell subsets.

• 出版日期2013-12-31