FISH Analysis of Sorted CD34+ Cells Cultured for 5 Days From Nonfrozen Maternal PB
| Sample . | Fetal Karyotype by CAAF . | No. of Cells Sorted . | No. of Cells Examined by FISH . | No. of Cells With XY Signal . | Hybridization Efficiency* X and Y Probe (%) . | Fetal Sex Correctly Assigned by FACS/FISH . |
|---|---|---|---|---|---|---|
| 32 | 46,XY | 2,527 | 11 | 0 | 71/76 | N |
| 33 | 46,XY | 4,467 | 3,064 | 5 | 63/85 | Y |
| 35 | 46,XY | 6,496 | 4,301 | 0 | 74/89 | N |
| 36 | 46,XY | 4,350 | 5,988 | 30 | 86/84 | Y |
| 37 | 46,XY | 1,695 | 2,039 | 11 | 74/95 | Y |
| 38 | 46,XY | 4,589 | 1,951 | 0 | 76/78 | N |
| 40 | 46,XY | 4,672 | 3,486 | 93 | 83/73 | Y |
| 34 | 46,XX | 1,729 | 1,505 | 0 | 88/86 | n/a |
| 39 | 46,XX | 3,248 | 1,677 | 2 | 89/78 | N |
| 41 | 46,XX | 3,631 | 1,895 | 5 | 68/80 | N |
| Sample . | Fetal Karyotype by CAAF . | No. of Cells Sorted . | No. of Cells Examined by FISH . | No. of Cells With XY Signal . | Hybridization Efficiency* X and Y Probe (%) . | Fetal Sex Correctly Assigned by FACS/FISH . |
|---|---|---|---|---|---|---|
| 32 | 46,XY | 2,527 | 11 | 0 | 71/76 | N |
| 33 | 46,XY | 4,467 | 3,064 | 5 | 63/85 | Y |
| 35 | 46,XY | 6,496 | 4,301 | 0 | 74/89 | N |
| 36 | 46,XY | 4,350 | 5,988 | 30 | 86/84 | Y |
| 37 | 46,XY | 1,695 | 2,039 | 11 | 74/95 | Y |
| 38 | 46,XY | 4,589 | 1,951 | 0 | 76/78 | N |
| 40 | 46,XY | 4,672 | 3,486 | 93 | 83/73 | Y |
| 34 | 46,XX | 1,729 | 1,505 | 0 | 88/86 | n/a |
| 39 | 46,XX | 3,248 | 1,677 | 2 | 89/78 | N |
| 41 | 46,XX | 3,631 | 1,895 | 5 | 68/80 | N |
PB (n = 10) was drawn at 10-13 weeks' gestation before any invasive diagnostic procedure and the FISH was performed without previous knowledge of the fetal karyotype. After sorting, cells were cultured for 5 days in serum-free media Epo, MGF, IL-6, GM-CSF/IL-3 fusion protein, M-CSF, and Flt-3 ligand (samples 40 and 41 only) and the results from both cultures were pooled.
Abbreviations: n/a; not applicable; CAAF, cytogenetic analysis of amniotic fluid.
Hybridization efficiency was controlled in each experiment using pure male and female mononuclear cells.