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. 2008 Aug 5;6(8):e191. doi: 10.1371/journal.pbio.0060191

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© 2008 Ahn et al.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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Sorted anti-LacZ TCR thymocytes were reaggregated with UEA-1–depleted CD45⁻ stromal cells from dGuo-treated fetal thymuses. (A) Flow cytometric profiles of reaggregate cultures initiated with 3.8 × 10⁵ DN + DP (1:10 ratio) and 5 × 10⁵ purified thymic stroma at day 5 (top) or day 6 (bottom). The sources of stromal cells and thymocytes are indicated at the top of the graphs. CD4 and CD8 profiles are shown along with the percentage of cells in each of the compartments. Typically, cell recoveries of the DN and DP thymocyte mixtures cultured with TDLacZ heterozygote (+/Δ) stroma were 20%–30% of thymocyte mixtures cultured with wild-type (+/+) stroma.

(B) A representative cell recovery when DN and DP cells were cultured separately with purified stroma from the wild type (+/+) versus the heterozygote (+/Δ ) mouse. Underlined numbers indicate the initial numbers of cells that were input.