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Demonstration of spontaneously dividing male fetal cells in maternal blood by negative magnetic cell sorting and fish |
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| Authors: | Yun-Ling Zheng PhD Sabrina D Craigo Cathy M Price Diana W Bianchi |
| Institution: | 1. Obstetrics and Gynecology, New England Medical Center, Tufts University School of Medicine, Boston, MA, U.S.A.;2. Leukaemia Research Fund Centre, Institute of Cancer Research, Chester Beatty Laboratories, London, U.K.;3. Departments of Pediatrics, New England Medical Center, Tufts University School of Medicine, Boston, MA, U.S.A.
Obstetrics and Gynecology, New England Medical Center, Tufts University School of Medicine, Boston, MA, U.S.A. |
| 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. |
| Keywords: | fetal cells in maternal blood fluorescence in situ hybridization magnetic-activated cell separation (MACS) CD45 |