Limiting dilution assays (LDAs) to determine the T-cell and B-cell differentiation potential of CD34+ cells.
(A) LDA design. Conditions for determining the potential of CD34+ cells to generate T cells a and B cells b are shown. For further details, see the materials and methods. (B, C) Cell cultures on D21 positive for T-cell precursors (B) defined as CD45RAhighCD7+CD5+CD1a+ cells, and B cells (C) defined as CD79aintra+ cells. (D) Analysis of the T-cell potential of CD34+ cells. Each point on the graph represents the mean value from three independent experiments. (E) The presence of T-cell precursors and B cells was assessed with the ELDA webtool, applying the maximum likelihood method to the Poisson model. Mean values (min-max) for three experiments are indicated for each group and each set of conditions. NS for P>0.05, **P<0.01, ***P<0.001.