High-efficiency retroviral gene transfer into murine high-proliferative-potential cells cycle-activated by cytosine arabinoside

We investigated cytosine arabinoside (Ara-C) as a potential agent for in vivo cycle activation of hematopoietic progenitors for the purpose of retroviral-mediated gene transfer. C57Bl mice were treated intraperitoneally with one of three regimens of Ara-C: a single 1,750 mg/kg dose (regimen 1), a 1,750 mg/kg dose on day 0, and a 1,500 mg/kg dose on day 2 (LD50) (regimen 2), or a 1,750 mg/kg dose on day 0 and a 1,500 mg/kg dose on day 3 (regimen 3). The high-proliferative-potential cells (HPPC)/10(5) cells were 47.0 +/- 7.5 pretreatment. The post-treatment HPPC cloning efficiencies were 40.6 +/- 3.4, 83.6 +/- 6.1, and 20.4 +/- 3.2 HPPC/10(5) cells on days 1, 2, and 4, respectively, with regimen 1; 60.0 +/- 7.9, 194.0 +/- 9.6, and 103.0 +/- 11.0 HPPC/10(5) cells 1, 2, and 4 days after the second Ara-C dose, respectively, with regimen 2; and 266 +/- 13.4, 132 +/- 23.9, and 118.0 +/- 5.7/10(5) cells 1, 2, and 4 days after the second Ara-C dose, respectively, with regimen 3. The transduction efficiency of HPPC from untreated animals with N2 viral supernatant was 4.9 +/- 5.8%.(ABSTRACT TRUNCATED AT 250 WORDS)