Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines

Hui-Yuen, Joyce; McAllister, Shane; Koganti, Siva; Hill, Erik; Bhaduri-McIntosh, Sumita

Temporarily store the mixes at 4°C in the dark. 3. In day three post-exposure to EBV, gently swirl flask to obtain a more uniform cell suspension. Remove 2ml from flask into a 2ml Eppendorf tube. Spin tube in a microcentrifuge at 3000 rpm at room temperature for 3 minutes. antibody. Plot CD19 + live B cells with fluorescence intensity for CD23 on x-axis and fluorescence intensity for CD58 on y-axis. Determine CD23 + and CD58 + cells by comparing to EBV-exposed cells stained with matched isotype control antibodies. Presence of CD23 hi CD58 + cells in EBV-exposed culture ( Figure 2C ) predicts successful outgrowth of EBV-infected growth transformed cells. In contrast, CD23 hi CD58 + cells do not emerge when cells are not exposed to EBV (Figure 2A ) or exposed to a non-immortalizing strain of EBV ( Figure 2B ). CD23 hi CD58 + cells have been experimentally demonstrated to undergo proliferation and subsequently establish LCL (14).