Table I.

Acquisition of CD3 mRNA by individual cells during DC development in culture from early T precursorsa

mRNA ProbedPercent Positive Cells
Thymocytes (control)CD4low precursors (not cultured)Cultured CD4low precursors
Day 1Day 2Day 3
β-Actin (control)99 ± 199 ± 198 ± 2100 ± 0100 ± 0
CD3ε97 ± 22 ± 390 ± 626 ± 232 ± 4
CD3δ85 ± 74 ± 694 ± 556 ± 200
CD3γ95 ± 53 ± 997 ± 1950
  • a Early prothymocytes (CD4low precursors) were purified from the adult mouse thymus and cultured with a mix of seven cytokines to induce the development of DC. Cells with DC morphology were evident by day 2 and no T lineage cells were detected. The cultured cells were harvested, the 30% dead cells removed, and the viable cells deposited on slides and processed for single cell in situ hybridization with the appropriate antisense probes. Where SDs are given, three to six individual samples were counted. Negative controls of cells with sense probes, or macrophages with antisense probes, always gave <5% positives.