Microsatellite analysis provides efficient confirmation of fetal trophoblast isolation from maternal circulation.
Fetal trophoblasts can be found in maternal circulation from an early stage of pregnancy and thus provide a potential source of DNA for non-invasive prenatal diagnosis. We have developed a two-step method for trophoblast isolation between the 8th and 12th week of pregnancy. Blood was sampled from 14 women undergoing termination of pregnancy or spontaneous abortion. Immunomagnetic beads precoated with HLA class I and II, and with anti-cytokeratin-18 monoclonal antibodies, were used to remove CD8+ and other maternal cells, and to select for fetal trophoblasts, respectively. Microsatellite analysis was performed on DNA extracted from the isolated, maternal, paternal and placental cells after PCR amplification. Recovery of the trophoblasts was confirmed in 13/14 cases (93%) by the identification of an identical microsatellite pattern for fetal and placental cells. Further evidence was the presence of heterozygous alleles of both maternal and paternal origin. The correct prediction of gender in all five male fetuses was an additional confirmation of trophoblast recovery. We conclude that trophoblasts can be effectively isolated from maternal blood in the first trimester, and by using polymorphic microsatellite markers to confirm sample purity, this method has potential future application in prenatal diagnosis.
Koumantaki
Y
Y
Department of Hygiene and Epidemiology, University of Athens, Medical School, Goudi, Athens, Greece.
Sifakis
S
S
Dragatis
G
G
Matalliotakis
I
I
Froudarakis
G
G
Papadopoulou
E
E
Koumantakis
E
E
D000293:Adolescent; D000328:Adult; D000911:Antibodies, Monoclonal; D002869:Chromosome Aberrations / Q000097:blood / Q000175:diagnosis*; D025063:Chromosome Disorders; D004247:DNA / Q000302:isolation & purification; D005260:Female; D005315:Fetal Diseases / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis / Q000379:methods / Q000592:standards*; D014327:Trophoblasts / Q000166:cytology*
D000911:Antibodies, Monoclonal; D004247:DNA