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CULTURE OF FETAL ERYTHROID PROGENITOR CELLS FROM MATERNAL BLOOD FOR NON‐INVASIVE PRENATAL GENETIC DIAGNOSIS
Author(s) -
VALERIO DOMENICO,
AIELLO RAFFAELE,
ALTIERI VINCENZO,
MALATO ANNA PAOLA,
FORTUNATO ADRIANA,
CANAZIO ANNA
Publication year - 1996
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/(sici)1097-0223(199612)16:12<1073::aid-pd38>3.0.co;2-d
Subject(s) - fetus , andrology , prenatal diagnosis , progenitor cell , biology , erythropoietin , fluorescence in situ hybridization , immunology , microbiology and biotechnology , pregnancy , medicine , stem cell , endocrinology , genetics , gene , chromosome
Fetal committed erythroid progenitors CFU‐E and M‐BFU‐E released into the maternal circulation during pregnancy are ideal candidates for in vitro proliferation since their lifespan is short and they can form colonies of 100–1000 cells in a semi‐solid medium. In order to propagate these cells with a high rate of purity, a strategy was devised based on their prior enrichment with biotin‐labelled human erythropoietin ligand and magnetic sorting before culturing in a suitable medium. Eight euploid pregnancies investigated in order to address this issue produced fetal clones in cultures with 18 per cent purity as assessed by polymerase chain reaction (PCR) analysis for Y‐specific sequences, immunocytochemical staining for fetal γ‐globin, and fluorescence in situ hybridization (FISH) study. The CFU‐E‐type colony was the most represented progenitor, followed by M‐BFU‐E, and only occasionally was the detection of CFU‐GEMM recorded. The retrospective diagnosis of two cases of fetal Down's syndrome by culturing fetal cells from maternal blood was accomplished for the first time. FISH analysis disclosed a strong presence of fetal trisomic cells (70 per cent and 40 per cent in the two cases). This strong presence would suggest a preferential leakage into maternal blood. The overall results of this study demonstrate that fetal cells can be cultured in vitro with reliable reproducibility, thus making the prospect of a non‐invasive prenatal genetic diagnosis realistic.