Objective: Efficient recovery of placental cells (and their subsequent characterisation) from the lower uterine pole (L.U.P) by trans-cervical flushing or aspiration.
Subjects: Women attending for termination of pregnancy (7-17 weeks' gestation) for social reasons. All patients gave their consent to the procedures outlined below.
Methods: Trans-cervical intrauterine flushing (using 0.15 M NaCl) or mucus aspiration. Embryo transfer catheters were used in both procedures. Fetal sexing was achieved by gene amplification of Y-specific DNA sequences (Y-PCR), and by in situ hybridisation (bright-field and fluorescence) to the Y-chromosome. Data were compared with results obtained from fetal tissues recovered following termination of pregnancy. Gender-independent tests for fetal cells utilised immunocytochemistry with trophoblast-specific antibodies and dual immunocytochemistry/ISH, where appropriate.
Results: (1) Fetal sexing by Y-PCR: 71/122 (58%) aspirates contained Y-specific DNA. In addition, the sexing of 72/86 (84%) aspirates and their corresponding samples of placental tissue, agreed exactly. (2) Microscopic detection of fetal cells. Placentally-derived syncytiotrophoblast was detected in 17/45 (38%) flushings and 39/173 (23%) aspirates. In most other Y-PCR+ samples which were negative for syncytiotrophoblast, Y-chromosome-bearing nuclei of unknown origin, were observed by ISH and immunocytochemical evidence for cytotrophoblastic cells was also uncovered.
Conclusions: Since Y-derived DNA can be detected in > 50% of flushings and aspirations, and gender-independent evidence for placental cells was obtained, regardless of fetal sex, we believe that most or all of these samples contained placental cells, including trophoblasts and naked nuclei. Trans-cervical placental cell recovery is a potentially valuable alternative to more invasive methods of aneuploid detection which require amniocentesis and CVS, provided its level of accuracy and above all, safety, can be evaluated.