Identification of embryo-fetal cells in celomic fluid using morphological and short-tandem repeats analysis.
OBJECTIVE
The main problem to wide acceptability of celocentesis as earlier prenatal diagnosis is contamination of the sample by maternal cells. The objective of this study was to investigate the cellular composition of celomic fluid for morphological discrimination between maternal and embryo-fetal cells.


METHODS
Celomic fluids were aspired by ultrasound-guided transcervical celocentesis at 7-9 weeks' gestation from singleton pregnancies before surgical termination for psychological reasons. DNA extracted from celomic fluid cells showed the same morphology, and quantitative fluorescent polymerase chain reaction (PCR) assay was performed to evaluate their fetal or maternal origin.


RESULTS
Six different types of non-hematological maternal and four different types of embryo-fetal cells were detected. The most common maternal cells were of epithelial origin. The majority of embryo-fetal cells were roundish with a nucleus located in an eccentric position near the wall. These cells were considered to be erythroblasts, probably derived from the yolk sac that serves as the initial site of erythropoiesis.


CONCLUSIONS
The combined use of morphology and DNA analysis makes it possible to select and isolate embryo-fetal cells, even when maternal contamination is high. This development provides the opportunity for the use of celocentesis for early prenatal diagnosis of genetic diseases and application of array comparative genomic hybridization. © 2016 John Wiley & Sons, Ltd.
Unit of Hematology for Rare Diseases of Blood and Blood-forming Organs, Regional Reference Laboratory for Screening Prenatal Diagnosis of Hemoglobinopathies, Palermo, Italy. a.giambona@villasofia.it.
Antonino
A
Giambona
Unit of Hematology for Rare Diseases of Blood and Blood-forming Organs, Regional Reference Laboratory for Screening Prenatal Diagnosis of Hemoglobinopathies, Palermo, Italy.
Filippo
F
Leto
Unit of Prenatal Diagnosis, Hospital Villa Sofia Cervello, Palermo, Italy.
Gianfranca
G
Damiani
Unit of Prenatal Diagnosis, Hospital Villa Sofia Cervello, Palermo, Italy.
Cristina
C
Jakil
Unit of Prenatal Diagnosis, Hospital Villa Sofia Cervello, Palermo, Italy.
Valentina
V
Cigna
Unit of Prenatal Diagnosis, Hospital Villa Sofia Cervello, Palermo, Italy.
Giovanna
G
Schillaci
Bionat Italia s.r.l., Palermo, Italy.
Giuseppe
G
Stampone
Reproductive Medicine Unit, ANDROS Day Surgery Clinic, Palermo, Italy.
Aldo
A
Volpes
Reproductive Medicine Unit, ANDROS Day Surgery Clinic, Palermo, Italy.
Adolfo
A
Allegra
Harris Birthright Research Centre for Fetal Medicine, King's College, London, UK.
Kypros H
KH
Nicolaides
Department of Obstetrics and Gynecology, Ioannina University Hospital, Ioannina, Greece.
George
G
Makrydimas
Unit of Hematology for Rare Diseases of Blood and Blood-forming Organs, Regional Reference Laboratory for Screening Prenatal Diagnosis of Hemoglobinopathies, Palermo, Italy.
Cristina
C
Passarello
Unit of Hematology for Rare Diseases of Blood and Blood-forming Organs, Regional Reference Laboratory for Screening Prenatal Diagnosis of Hemoglobinopathies, Palermo, Italy.
Aurelio
A
Maggio
D001826:Body Fluids / Q000166:cytology*; D055028:Comparative Genomic Hybridization; D004247:DNA / Q000032:analysis*; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005333:Fetus / Q000166:cytology*; D005453:Fluorescence; D005865:Gestational Age; D058746:Gestational Sac; D006801:Humans; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis
D004247:DNA