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Detection of fetal erythroid cells from maternal blood using fluorescence in situ hybridization and liquid culture.
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MedLine Citation:
PMID:  11306738     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 0.1 x 10(6)-1.0 x 10(6)/mL with a purity of 84.0-97.3%. Hb F stain showed total erythroid cells of approximately 0.4 x 10(4)-9.8 x 10(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.
Authors:
J Y Han; K H Kim; J I Park; I H Kim; G H Je
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of Korean medical science     Volume:  16     ISSN:  1011-8934     ISO Abbreviation:  J. Korean Med. Sci.     Publication Date:  2001 Apr 
Date Detail:
Created Date:  2001-04-18     Completed Date:  2001-07-19     Revised Date:  2011-03-16    
Medline Journal Info:
Nlm Unique ID:  8703518     Medline TA:  J Korean Med Sci     Country:  Korea (South)    
Other Details:
Languages:  eng     Pagination:  145-9     Citation Subset:  IM    
Affiliation:
Department of Clinical Pathology, Dong-A University College of Medicine, Pusan, Korea. jyhan@daunet.donga.ac.kr
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MeSH Terms
Descriptor/Qualifier:
Cell Culture Techniques / methods
Cell Separation / methods*
Chromosome Aberrations / diagnosis
Chromosome Disorders
Erythroblasts*
Female
Genetic Testing
Humans
In Situ Hybridization, Fluorescence*
Karyotyping
Maternal-Fetal Exchange*
Pregnancy
Prenatal Diagnosis / methods*

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Full Text
Journal Information
Journal ID (nlm-ta): J Korean Med Sci
Journal ID (pmc): jkms
ISSN: 1011-8934
ISSN: 1598-6357
Publisher: Korean Academy of Medical Sciences
Article Information
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Print publication date: Month: 4 Year: 2001
Volume: 16 Issue: 2
First Page: 145 Last Page: 149
ID: 3054725
PubMed Id: 11306738
Publisher Item Identifier: 200104145

Detection of fetal erythroid cells from maternal blood using fluorescence in situ hybridization and liquid culture.
J. Y. Han Email: jyhan@daunet.donga.ac.kr
K. H. Kim
J. I. Park
I. H. Kim
G. H. Je
Department of Clinical Pathology, Dong-A University College of Medicine, Pusan, Korea.


Article Categories:
  • Research Article


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