Comparison of MNC and BAMP screening for healthy volunteers and a patient with MDS.

(A) MNCs isolated from the PB of healthy volunteers were subjected to anti-AC133 affinity chromatography, followed by fluorescence-activated cell sorting with anti-AC133. The eluted (AC133⁺) cells and the original MNCs were analyzed for surface expression of CD34 and CD38 by flow cytometry (upper panels). The percentage of CD34⁺cells in each sample is indicated. Wright-Giemsa staining of each cell preparation is also shown (lower panels, original magnification ×100). (B) Purification of AC133⁺ cells from a patient with MDS-associated leukemia. MNCs isolated from the PB of the patient (ID no. 7) were subjected to anti-AC133 affinity chromatography. The MNCs and AC133⁺ cells were then analyzed for surface expression of CD34 and CD38 and subjected to Wright-Giemsa staining as described for panel A, original magnification ×100. (C) Total RNA isolated from the MNCs of healthy volunteers and the patient with MDS was used to synthesize cDNA labeled with Cy5 and Cy3 dyes, respectively. A mixture of the labeled cDNA preparations was then subjected to hybridization with a cDNA microarray containing fragments of 382 cancer-related genes (upper panel). Red and green spots thus indicate genes specifically expressed in the healthy volunteers or in the patient, respectively; yellow spots correspond to genes expressed to similar extents in both samples. RNA prepared from purified AC133⁺ cells of the healthy volunteers and the patient was similarly analyzed (lower panel). A part of the scanned image is demonstrated. Some spots, including those numbered, exhibited opposite patterns of expression by MNC screening and BAMP screening.