The use of non-physiological conditions to isolate fetal cells from maternal blood.
Fetal cells are always present in maternal blood starting in the first trimester of pregnancy, however a rapid, simple, and consistent procedure for their isolation for prenatal non-invasive genetic investigation is still lacking. Sensitivity and recovery of fetal cells is jeopardized by the minute amount of circulating fetal cells and their loss during the enrichment procedure. We report here a single-step approach to isolate fetal cells from maternal blood which relies on the use of non-physiological conditions to modify cell densities before their separation in a density gradient and in a newly developed cell separation device. Isolated fetal cells have been investigated using cytochemistry, Soret band absorption microscopy, monoclonal antibodies for epsilon- and gamma-chain-Hb, monoclonal antibody for i-antigen, and by fluorescence in situ hybridization (FISH). Fetal cells were always detected in all 105 maternal blood samples investigated and fetal aneuploidies were correctly diagnosed by FISH, in a pilot study of pathological pregnancies, in fetal cells isolated from maternal blood obtained either before or after invasive procedure.
Sitar
Giammaria
G
Department of Medicine Policlinico San Matteo and University of Pavia, Italy. g.sitar@smatteo.pv.it
Brambati
Bruno
B
Baldi
Marina
M
Montanari
Laura
L
Vincitorio
Massimo
M
Tului
Lucia
L
Forabosco
Antonino
A
Ascari
Edoardo
E
D000782:Aneuploidy; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001773:Blood Cells / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005315:Fetal Diseases / Q000473:pathology; D005319:Fetal Hemoglobin / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D010865:Pilot Projects; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis / Q000379:methods*
D000911:Antibodies, Monoclonal; D005319:Fetal Hemoglobin