Characterization of first trimester fetal erythroblasts for non-invasive prenatal diagnosis

Choolani, Mahesh, O’Donoghue, Keelin, Talbert, David, Kumar, Sailesh, Roberts, Irene, Letsky, Elizabeth, Bennett, Phillip R. and Fisk , Nicholas M. (2003) Characterization of first trimester fetal erythroblasts for non-invasive prenatal diagnosis. Molecular Human Reproduction, 9 4: 227-235. doi:10.1093/molehr/gag027


Author Choolani, Mahesh
O’Donoghue, Keelin
Talbert, David
Kumar, Sailesh
Roberts, Irene
Letsky, Elizabeth
Bennett, Phillip R.
Fisk , Nicholas M.
Title Characterization of first trimester fetal erythroblasts for non-invasive prenatal diagnosis
Journal name Molecular Human Reproduction   Check publisher's open access policy
ISSN 1360-9947
1460-2407
Publication date 2003-04-01
Year available 2003
Sub-type Article (original research)
DOI 10.1093/molehr/gag027
Open Access Status DOI
Volume 9
Issue 4
Start page 227
End page 235
Total pages 9
Place of publication Oxford, U.K.
Publisher Oxford University Press
Language eng
Subject 111401 Foetal Development and Medicine
111402 Obstetrics and Gynaecology
1114 Paediatrics and Reproductive Medicine
Abstract Isolating fetal erythroblasts from first trimester maternal blood offers a promising non-invasive alternative for prenatal diagnosis. The aim of this study was to characterize the biological properties of first trimester primitive erythroblasts to facilitate their enrichment from first trimester maternal blood. Primitive erythroblasts were the predominant cell type until 12 weeks gestation, after which time their numbers declined steeply; 100% were epsilon-globin-positive versus <0.06% definitive erythroblasts. Buoyant densities of first trimester fetal erythroblasts ranged from 1.077 to 1.130 g/ml, and optimal recoveries were obtained with Percoll 1118. Although primitive erythroblasts carried a negative surface charge and were resistant to NH4Cl lysis, these properties had only a limited role in fetal cell enrichment. Immunophenotyping showed that primitive, like definitive, erythroblasts were GPA+, CD47+, CD45- and CD35-, whereas CD71 expression was weak/undetectable on primitive erythroblasts but strongly positive on 100% of definitive erythroblasts; primitive erythroblasts were also CD36-whereas definitive erythroblasts were CD36+. We therefore used CD45/GPA selection of Percoll 1118-separated cells to demonstrate successful enrichment of male ε-globin-positive fetal erythroblasts from model mixtures, and as proof of principle from some first trimester maternal blood samples. Fetal cell enrichment protocols based on first trimester ε-globin-positive primitive erythroblasts may allow reliable enrichment of fetal cells from maternal blood for early non-invasive prenatal diagnosis of genetic disorders.
Formatted abstract
Isolating fetal erythroblasts from first trimester maternal blood offers a promising non-invasive alternative for prenatal diagnosis. The aim of this study was to characterize the biological properties of first trimester primitive erythroblasts to facilitate their enrichment from first trimester maternal blood. Primitive erythroblasts were the predominant cell type until 12 weeks gestation, after which time their numbers declined steeply; 100% were ε-globin-positive versus <0.06% de®nitive erythroblasts. Buoyant densities of first trimester fetal erythroblasts ranged from 1.077 to 1.130 g/ml, and optimal recoveries were obtained with Percoll 1118. Although primitive erythroblasts carried a negative surface charge and were resistant to NH4Cl lysis, these properties had only a limited role in fetal cell enrichment. Immunophenotyping showed that primitive, like definitive, erythroblasts were GPA+, CD47+, CD45± and CD35±, whereas CD71 expression was weak/undetectable on primitive erythroblasts but strongly positive on 100% of definitive erythroblasts; primitive erythroblasts were also CD36± whereas definitive erythroblasts were CD36+. We therefore used CD45/GPA selection of Percoll 1118-separated cells to demonstrate successful enrichment of male ε-globin-positive fetal erythroblasts from model mixtures, and as proof of principle from some first trimester maternal blood samples. Fetal cell enrichment protocols based on first trimester ε-globin-positive primitive erythroblasts may allow reliable enrichment of fetal cells from maternal blood for early non-invasive prenatal diagnosis of genetic disorders.
© 2003 European Society of Human Reproduction and Embryology.

Keyword ε-globin
Erythroblasts
Fetal cells
maternal blood
Prenatal diagnosis
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Medicine Publications
 
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Created: Thu, 19 Mar 2009, 00:53:20 EST by Maryanne Watson on behalf of Faculty Of Health Sciences