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Frequency of fetal cells in sorted subpopulations of nucleated erythroid
and CD34+ hematopoietic progenitor cells from maternal peripheral blood
MT Little, S Langlois, RD Wilson and PM Lansdorp
Department of Medical Genetics, University of British Columbia, Vancouver,
Canada.
Fetal cells that circulate in maternal peripheral blood (PB) during
pregnancy offer a potential source of nucleated fetal material for
noninvasive prenatal diagnosis. Fluorescence-activated cell sorting was
used to target two populations of fetal cells: nucleated erythroid cells
(NECs; CD71/glycophorin-A+ CD45(lo-int) CD34-) and hematopoietic progenitor
cells (CD34+ cells; CD34++ CD71/glycophorin-A- CD45(int)). Fetal cells were
detected by fluorescence in situ hybridization (FISH) using directly
conjugated chromosome X and Y probes in 65% (13 of 20) of the maternal PBs
(fetal karyotype 46,XY). The frequency of fetal cells isolated from the NEC
and CD34+ fractions was, respectively, 0 to 14 and 0 to 7 cells per 2 x
10(7) previously frozen maternal cells (approximately 20 mL of blood). In
nonfrozen samples, the yield and recovery of fetal cells was moderately
improved. Culturing the CD34+ sorted fractions in serum-free media with
cytokines improved the quality of the FISH preparations and resulted in a
slight expansion in detectable fetal cells. The frequency of fetal cells
isolated from cultured CD34+ fractions was 0 to 35 and 0 to 93 cells per 2
x 10(7) previously frozen and nonfrozen maternal PB cells, respectively.
These results document the isolation, characterization, and enumeration of
fetal cells from the maternal periphery that appear to be present in most,
but not all, samples analyzed.
Volume 89,
Issue 7,
pp. 2347-2358,
04/01/1997
Copyright © 1997 by The American Society of Hematology
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