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Detection of fetal erythroid cells from maternal blood using fluorescence in situ hybridization and liquid culture
Journal of Korean Medical Science ; : 145-149, 2001.
Artigo em Inglês | WPRIM | ID: wpr-179360
ABSTRACT
Fetal nucleated erythrocytes circulating in maternal blood are a potential source of fetal DNA for noninvasive prenatal genetic diagnosis. However, the estimated ratio of fetal to maternal cells is extremely small. In order to enrich these cells, we performed direct culture using a two-phase liquid system. Mononuclear cells were obtained from maternal blood samples at 8-10(+3) weeks of gestation and cultured in the first phase. After 4-5 days, the nonadherent cells were harvested and recultured with erythropoietin in the second phase for another 3-5 days. We examined cellular morphology, and counted the number of benzidine- positive cells and the percentage of glycophorin A/CD71 positive erythroid cells. We also did Kleihauer-Betke stain for Hb F, polymerase chain reaction (PCR) for SRY/DYZ1, chromosome analysis, and fluorescence in situ hybridization (FISH). The number of total erythroid cells reached about 0.1x10(6)-1.0x10(6)/mL with a purity of 84.0-97.3%. Hb F stain showed total erythroid cells of approximately 0.4x10(4)-9.8x10(4)/mL. Male DNA was detected in one case by PCR. In this case, the XY karyotype was confirmed by FISH and amniocentesis. This approach provides enriched source of fetal cells for further prenatal genetic analysis without complicated separation or sorting procedures.
Assuntos

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Separação Celular / Eritroblastos / Testes Genéticos / Aberrações Cromossômicas / Hibridização in Situ Fluorescente / Técnicas de Cultura de Células / Cariotipagem / Troca Materno-Fetal Tipo de estudo: Estudo diagnóstico / Estudo prognóstico Limite: Feminino / Humanos / Gravidez Idioma: Inglês Revista: Journal of Korean Medical Science Ano de publicação: 2001 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Separação Celular / Eritroblastos / Testes Genéticos / Aberrações Cromossômicas / Hibridização in Situ Fluorescente / Técnicas de Cultura de Células / Cariotipagem / Troca Materno-Fetal Tipo de estudo: Estudo diagnóstico / Estudo prognóstico Limite: Feminino / Humanos / Gravidez Idioma: Inglês Revista: Journal of Korean Medical Science Ano de publicação: 2001 Tipo de documento: Artigo