Tracking OT-I T Cell Migration

OT-I CD8⁺ T cells in suspension were stained with 2.5 μM Cell Tracker Violet BMQC Dye (C10094, Invitrogen) for 30 min. CM from tumor cells (0.75 mL of medium containing 5% FBS) were plated in the lower wells of a chamber assay plate, and OT-I cells (2.5 × 10⁵ cells in 0.1 mL of medium containing 1% FBS) were placed in the upper wells of the insert. After co-incubation for 1, 2, 4, or 6 h, cells were collected from the bottom wells and subjected to flow cytometry to identify BMQC⁺ cells^{70 (link)}.

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Yan J., Zhao Q., Gabrusiewicz K., Kong L.Y., Xia X., Wang J., Ott M., Xu J., Davis R.E., Huo L., Rao G., Sun S.C., Watowich S.S., Heimberger A.B, & Li S. (2019). FGL2 promotes tumor progression in the CNS by suppressing CD103+ dendritic cell differentiation. Nature Communications, 10, 448.

Publication 2019

Cell Tracker Violet BMQC Dye

Assay Cd8 t cells Cells Dye 2 Flow cytometry Tumor Violet

Corresponding Organization :

Other organizations : Capital Medical University, The University of Texas MD Anderson Cancer Center

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Variable analysis

independent variables

Co-incubation time (1, 2, 4, or 6 hours)

dependent variables

Percentage of BMQC+ cells identified by flow cytometry

control variables

OT-I CD8+ T cells in suspension
Cell Tracker Violet BMQC Dye concentration (2.5 μM)
Cell Tracker Violet BMQC Dye incubation time (30 minutes)
Tumor cell-conditioned medium (0.75 mL of medium containing 5% FBS)
Number of OT-I cells (2.5 × 10^5 cells in 0.1 mL of medium containing 1% FBS)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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