Do fetal extravillous trophoblasts circulate in maternal blood postpartum

INTRODUCTION: Circulating fetal extravillous trophoblasts may offer a superior alternative to cell-free fetal DNA for noninvasive prenatal testing. Cells of fetal origin are a pure source of fetal genome, hence, unlike cell-free noninvasive prenatal test, fetal cell-based noninvasive prenatal test is not expected to be affected by maternal DNA. However, circulating fetal cells from previous pregnancies may lead to confounding results. MATERIAL AND METHODS: In order to study whether fetal trophoblast cells persist in maternal circulation postpartum, blood samples were collected from 11 women who had given birth to a boy, with blood sampling at 1-3 days (W0), 4-5 weeks (W4-5), around 8 weeks (W8), and around 12 weeks (W12) postpartum. The existence of fetal extravillous trophoblasts was verified either by X and Y chromosome fluorescence in-situ hybridization analysis, or by short tandem repeat analysis. In order to exclude technological bias in isolating fetal cells, blood samples were also collected from 10 pregnant women in gestational age of 10-14 weeks, the optimal time frame for cell-based noninvasive prenatal test sampling. All the samples were processed according to protocols established by ARCEDI Biotech (Vejle, Denmark) for fetal extravillous trophoblasts enrichment and isolation. RESULTS: Fetal extravillous trophoblasts were found in all the 10 samples from pregnant women in gestational age of 10-14 weeks. However, only four out of 11 blood samples taken from women, one to three days postpartum rendered fetal extravillous trophoblasts. And only two out of 11 samples rendered fetal extravillous trophoblasts at four weeks postpartum. CONCLUSIONS: In this preliminary dataset on few pregnancies, none of the samples rendered any fetal cells at or after eight weeks postpartum, showing that cell-based noninvasive prenatal testing based on fetal extravillous trophoblasts is unlikely to be influenced by circulating cells from previous pregnancies.