Abstract

<p>CD34<sup>+</sup> cells purified from cord blood were transduced with void UMG-LV6 vector or UMG-LV6 carrying the MLL-AF9 cDNA (UMG-LV6-MA). (<b>A</b>) FACS analysis of CD34<sup>+</sup> cells 5 days after transduction. The percentages of EGFP positive cells are indicated. (<b>B</b>) Q-RT-PCR analysis of the expression of MLL-AF9 in CD34<sup>+</sup> cells transduced with UMG-LV6-MA. The expression level was compared to that of the MLL-AF9-positive THP-1 cells, assumed as 1. (<b>C</b>) 1×10<sup>4</sup> CD34<sup>+</sup> cells transduced with void UMG-LV6 vector or with UMG-LV6-MA/well were plated in triplicate in 6-well plates in cytokine-driven cultures in the presence of 100 ng/ml of stem cell factor, FLT3 ligand and thrombopoietin. The culture medium was refreshed weekly, and the cell numbers were determined two weeks after plating. (<b>D</b>) The number of clonogenic progenitors in CD34<sup>+</sup> cells transduced with void UMG-LV6 vector or with UMG-LV6-MA after two weeks of cytokine-driven culture was determined by clonogenic assays in methylcellulose as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114795#s2" target="_blank">Matherials and Methods</a>.</p

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