Abstract
PurposeThe aim of this study was to determine if pregnancy-associated plasma protein-A (PAPP-A), typically measured in maternal serum and a potential predictor of adverse maternal and fetal outcomes such as spontaneous miscarriage, pre-eclampsia, and stillbirth, is expressed in blastocoel fluid–conditioned media (BFCM) at the embryonic blastocyst stage.DesignThis is an in vitro study.MethodsBFCM samples from trophectoderm-tested euploid blastocysts (n = 80) from in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) patients were analyzed for PAPP-A mRNA. BFCM was obtained from blastocyst stage embryos in 20 uL drops. Blastocysts underwent trophectoderm biopsy for preimplantation genetic testing for aneuploidy prior to blastocyst vitrification and BFCM collection for snap freezing. cfDNA was synthesized using BFCM collected from 80 individual euploid blastocysts. Next, real-time qPCR was performed to detect expression of PAPP-A with GAPDH for normalization of expression in each sample.ResultsPAPP-A mRNA was detected in 45 of 80 BFCM samples (56.3%), with varying levels of expression across samples.ConclusionOur study demonstrates the expression of PAPP-A in BFCM. To our knowledge, this is the first study to report detection of PAPP-A mRNA in BFCM. Further studies are required and underway to investigate a greater number of BFCM samples as well as the possible correlation of PAPP-A expression with pregnancy outcomes of transferred euploid blastocysts. If found to predict IVF and obstetric outcomes, PAPP-A may provide additional information along with embryonic euploidy for the selection of the optimal blastocyst for embryo transfer.
Highlights
There is a great need for early detection and prevention of adverse maternal and fetal prenatal outcomes
Since the time that it was first detected in the serum plasma of pregnant women [11], maternal serum Pregnancyassociated plasma protein A (PAPP-A) has been the subject of many publications, and investigators have studied the secretion of PAPP-A by trophoblastic cells as well [1, 12]
Of the 45 blastocysts that had PAPP-A mRNA detected in their blastocoel fluid–conditioned media (BFCM) samples, 36 blastocysts (80%) had trophectoderm grading of A and 9 blastocysts had trophectoderm grading of B; of the 35 blastocysts that had no detectable PAPP-A mRNA in their BFCM samples, 28 (80%) had trophectoderm grading of A and 7 blastocysts had trophectoderm grading of B
Summary
There is a great need for early detection and prevention of adverse maternal and fetal prenatal outcomes. Pregnancyassociated plasma protein A (PAPP-A), predominantly produced by placental trophoblastic cells [1] and measured in maternal serum during the first trimester for clinical purposes, has been shown to be a potential predictor of adverse obstetric outcomes [2,3,4,5,6] such as spontaneous miscarriage, small for gestational age (SGA)/intrauterine growth restriction (IUGR), pre-eclampsia, and intrauterine fetal demise (IUFD) [1, 7]. Recent data suggest that low levels of PAPPA in maternal serum is associated with abnormal placentation which may explain potential downstream maternal and fetal complications [13]. Existing data indicate that PAPP-A is expressed at the level of the human ovary [15], in granulosa cells, theca cells [16, 17], follicular fluid [10], and in cumulus granulosa cell masses [18], as well as downstream in trophoblastic cells, there is lack of data regarding the potential expression of PAPP-A at the human embryonic level in vitro
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