title: Demonstration of spontaneously dividing male fetal cells in maternal blood by negative magnetic cell sorting and fish.
abstract: We investigated a case of massive feto-maternal bleeding by using negative magnetic cell sorting (MACS) and fluorescent in situ hybridization (FISH). A 37-year-old pregnant woman had an uncomplicated amniocentesis for advanced maternal age at 16 weeks' gestation. The fetal karyotype was 46,XY. At 19 weeks' gestation, she had a minor car accident and slight vaginal bleeding. A subsequent Kleihauer-Betke test showed a 140 ml feto-maternal haemorrhage. Serial sonographic examinations indicated a normal fetus and placenta. We performed FISH analysis on maternal peripheral blood at 25 weeks. Anti-CD45 and MACS were used to deplete maternal leucocytes, enriching the proportion of fetal nucleated erythrocytes present. The isolated cells were analysed by using dual-colour FISH with X and Y specific probes. Approximately 65,800 nucleated cells were obtained after MACS depletion. A total of 234 cells were analysed by FISH. The results revealed that 70 of the nucleated cells (30 per cent) were male with one X and one Y signal. Among these cells, six male metaphases were observed in spontaneously dividing cells.
authors.affiliation: Department of Pediatrics, New England Medical Center, Tufts University School of Medicine, Boston, MA, USA.
authors.forename: Y L
authors.initials: YL
authors.lastname: Zheng
authors.forename: S D
authors.initials: SD
authors.lastname: Craigo
authors.forename: C M
authors.initials: CM
authors.lastname: Price
authors.forename: D W
authors.initials: DW
authors.lastname: Bianchi
mesh_terms: D000328:Adult; D001773:Blood Cells / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005331:Fetomaternal Transfusion / Q000175:diagnosis*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008280:Magnetics; D008297:Male; D011247:Pregnancy; D014960:X Chromosome; D014998:Y Chromosome