A method for high purity sorting of rare cell subsets applied to T_DC

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• 作者：Mirela Kuka ; Jonathan D. Ashwell
• 关键词：Flow cytometry ; Sorting ; TDC ; Purification
• 刊名：Journal of Immunological Methods
• 出版年：31 December, 2013
• 年：2013
• 卷：400-401
• 期：Complete
• 页码：111-116
• 全文大小：1082 K

文摘

T_DC are a recently described subset of polyclonal 伪尾 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.