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Enrichment and detection of fetal erythroid cells from maternal peripheral blood using liquid culture
Author(s) -
Han JinYeong,
Lee YoungHee,
Sin SangDong,
Park JooIn,
Kim InHoo,
Je GooHwa,
Rodgers Griffin P.
Publication year - 2001
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 - 10.1002/1097-0223(200101)21:1<22::aid-pd987>3.0.co;2-d
Subject(s) - fetus , amniocentesis , andrology , testis determining factor , biology , prenatal diagnosis , peripheral blood mononuclear cell , erythropoietin , microbiology and biotechnology , karyotype , immunology , pregnancy , medicine , endocrinology , y chromosome , genetics , chromosome , gene , in vitro
Isolating fetal cells from maternal blood for prenatal genetic analysis is the least invasive method currently being investigated. In order to enrich these fetal erythroid cells we employed a two‐phase liquid culture system that supports the growth and differentiation of erythroid progenitor cells. Mononuclear cells were separated from ten maternal blood samples at 8–14 +3 weeks of gestation and cultured in the first phase. After 4–5 days, the non‐adherent cells were harvested and recultured with erythropoietin for another 3–4 days. The mean number of total erythroid cells reached approximately 0.77×10 6 /ml with an average purity of 90.5%. Hb F stain disclosed fetal erythroid cells averaging ∼5.7%, therefore the mean number of fetal erythroid cells isolated was 0.49×10 5 /ml. Female karyotypes were only observed while counting 5–66 metaphases. However, male DNA (SRY or DYZ1) was detected by PCR in 7/10 cases; in four of these cases and in one SRY‐negative pregnancy the 46,XY karyotype was found by amniocentesis performed during the second trimester. This liquid culture system provided an enriched source of fetal cells without the need for more complicated separation or sorting procedures. Copyright © 2001 John Wiley & Sons, Ltd.