Detection of fetal erythroid cells from maternal blood using fluorescence in situ hybridization and liquid culture.
10.3346/jkms.2001.16.2.145
- Author:
Jin Yeong HAN
1
;
Kyeong Hee KIM
;
Joo In PARK
;
In Hoo KIM
;
Goo Hwa JE
Author Information
1. Department of Clinical Pathology, Dong-A University College of Medicine, Pusan, Korea. jyhan@daunet.donga.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Fetal Blood;
Liquid Culture;
Material Blood;
Kleihauer-Betke Stain;
Glycophorin A/CD71;
In Situ Hybridization;
Prenatal Diagnosis
- MeSH:
Cell Culture/methods;
Cell Separation/*methods;
Chromosome Abnormalities/diagnosis;
*Erythroblasts;
Female;
Genetic Screening;
Human;
*In Situ Hybridization, Fluorescence;
Karyotyping;
*Maternal-Fetal Exchange;
Pregnancy;
sPrenatal Diagnosis/*methods
- From:Journal of Korean Medical Science
2001;16(2):145-149
- CountryRepublic of Korea
- Language:English
-
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.
