Phenotyping EBV-Infected Cell Lines Using Flow Cytometry

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Resumo

Source: Xu, C., et al. An Efficient and Simple Method to Establish NK and T Cell Lines from Patients with Chronic Active Epstein-Barr Virus Infection. J. Vis. Exp. (2018).

This video demonstrates the phenotyping of EBV-infected T and NK cell lines using flow cytometry, highlighting the preparation, antibody staining, and analysis steps to identify lineage-specific surface markers. It shows how marker profiles confirm distinct T-cell and NK-cell populations.

Protocolo

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. Cell Phenotyping by Flow Cytometry

When cells have expanded, collect 1×10⁶ cells to determine the phenotypes of cell lines. Centrifuge cells at 240 x g for 5 min at room temperature (NK/T cells will precipitate at the bottom of the tube).
Discard the supernatant, wash the precipitation by adding 7 mL PBS. Centrifuge for 5 min at 240 x g at room temperature. Repeat once.
Add 200 µL human serum to each tube, mix well and incubate for 10 min at room temperature.
Centrifuge cells a....

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Resultados

Figure 1: Representative gating strategy for flow cytometry analysis of cell lines L196 and M296. PBMC were used for adjusting FSC and SSC voltages. The cluster of live and single lymphocytes was gated for P1 in the FSC and SSC plot. The i.......