Enrichment and identification of fetal trophoblast cells from first trimester maternal cervical lavage and uterine blood specimens.
First trimester prenatal diagnosis of fetal aneuploidies is an active area of research despite years of disappointing data employing maternal peripheral blood samples. To remedy this situation we have investigated other first trimester maternal specimens attempting to find a consistent fetal cell source. Using our previously established positive enrichment procedure along with a commercially available depletion method, fetal trophoblast cells were identified employing immunocytochemistry using an antibody cocktail or by using mRNA in-situ hybridization employing a cocktail of trophoblast specific probes. Fetal origin of positively identified cells was verified using interphase fluorescent in-situ hybridization (FISH) for X and Y-chromosomes. Artificial model systems were established that indicated yields of trophoblast cells and allowed the enrichment procedure to be optimized for minimal losses from maternal specimens. We demonstrate herein that blood drawn from maternal vessels near the placental implantation site to be the most consistent source of fetal cells from any first trimester maternal specimen described to date. In addition, a high yield of multinucleated syncytiotrophoblast cells was obtained using a cell depletion strategy to enrich the target cells. The safety of the procedure or even the clinical utility of blood drawn from maternal vessels near the placental implantation site is yet to be demonstrated.
Schueler
P A
PA
Chief Technology Office, Roche Diagnostics, Berkeley, CA, USA.
Yamanishi
D T
DT
Collarini
E J
EJ
Pearson
J
J
Lee
Y
Y
Cain
C A
CA
Edwards
J
J
Foltz
L
L
Wu
X
X
Mahoney
W
W
D002469:Cell Separation; D002584:Cervix Uteri / Q000166:cytology*; D002869:Chromosome Aberrations; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D017404:In Situ Hybridization, Fluorescence; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis; D012333:RNA, Messenger / Q000032:analysis; D007507:Therapeutic Irrigation; D014327:Trophoblasts / Q000166:cytology*; D014599:Uterus / Q000098:blood supply*; D014960:X Chromosome; D014998:Y Chromosome
D012333:RNA, Messenger