Figure 5.

Long-term cultures of CB CD34⁺ cells transduced with STAT5A(1*6) on MS5 stroma. (A) CB CD34⁺ cells were transduced with STAT5A(1*6) and plated on MS5 to generate CAFCs, which were harvested and replated onto new MS5 at day 10, weeks 3, 5, and 7 (not depicted), and weeks 10, 13, and 15 (indicated by the arrows). Cultures were weekly demidepopulated and cell counts of a representative experiment out of two independent experiments are shown. At week 14, nonadherent cells were used for FACS analysis (see Table I), RT-PCR analysis (B), in which total RNA was isolated, and used in RT-PCR reactions with primers that amplify the STAT5A(1*6)-IRES2 cassette, cytospins (C), or CFC assays (D), in which cells were plated in methylcellulose.