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

被引:44
作者
Choolani, M [1 ]
O'Donoghue, K [1 ]
Talbert, D [1 ]
Kumar, S [1 ]
Roberts, I [1 ]
Letsky, E [1 ]
Bennett, PR [1 ]
Fisk, NM [1 ]
机构
[1] Univ London Imperial Coll Sci Technol & Med, Queen Charlottes & Chelsea Hosp, Div Paediat Obstet & Gynaecol, Inst Reprod & Dev Biol,Wolfson & Weston Res Ctr F, London W12 0NN, England
关键词
E-globin; erythroblasts; fetal cells; maternal blood; prenatal diagnosis;
D O I
10.1093/molehr/gag027
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 epsilon-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 epsilon-globin-positive primitive erythroblasts may allow reliable enrichment of fetal cells from maternal blood for early non-invasive prenatal diagnosis of genetic disorders.
引用
收藏
页码:227 / 235
页数:9
相关论文
共 66 条
  • [1] Al-Mufti R, 2001, HAEMATOLOGICA, V86, P357
  • [2] ALTER BP, 1979, BLOOD, V53, P279
  • [3] ONE-STEP ENRICHMENT OF NUCLEATED RED-BLOOD-CELLS - A POTENTIAL APPLICATION IN PERINATAL DIAGNOSIS
    BHAT, NM
    BIEBER, MM
    TENG, NNH
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 1993, 158 (02) : 277 - 280
  • [4] ONE-STEP SEPARATION OF HUMAN FETAL LYMPHOCYTES FROM NUCLEATED RED-BLOOD-CELLS
    BHAT, NM
    BIEBER, MM
    TENG, NNH
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 1990, 131 (01) : 147 - 149
  • [5] Significant fetal-maternal hemorrhage after termination of pregnancy: Implications for development of fetal cell microchimerism
    Bianchi, DW
    Farina, A
    Weber, W
    Delli-Bovi, LC
    DeRiso, M
    Williams, JM
    Klinger, KW
    [J]. AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY, 2001, 184 (04) : 703 - 706
  • [6] ERYTHROID-SPECIFIC ANTIBODIES ENHANCE DETECTION OF FETAL NUCLEATED ERYTHROCYTES IN MATERNAL BLOOD
    BIANCHI, DW
    ZICKWOLF, GK
    YIH, MC
    FLINT, AF
    GEIFMAN, OH
    ERIKSON, MS
    WILLIAMS, JM
    [J]. PRENATAL DIAGNOSIS, 1993, 13 (04) : 293 - 300
  • [7] FETAL CELLS IN MATERNAL BLOOD - DETERMINATION OF PURITY AND YIELD BY QUANTITATIVE POLYMERASE CHAIN-REACTION
    BIANCHI, DW
    SHUBER, AP
    DEMARIA, MA
    FOUGNER, AC
    KLINGER, KW
    [J]. AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY, 1994, 171 (04) : 922 - 926
  • [8] Fetal gender and aneuploidy detection using fetal cells in maternal blood: analysis of NIFTY I data
    Bianchi, DW
    Simpson, JL
    Jackson, LG
    Elias, S
    Holzgreve, W
    Evans, MI
    Dukes, KA
    Sullivan, LM
    Klinger, KW
    Bischoff, FZ
    Hahn, S
    Johnson, KL
    Lewis, D
    Wapner, RJ
    [J]. PRENATAL DIAGNOSIS, 2002, 22 (07) : 609 - 615
  • [9] ISOLATION OF FETAL DNA FROM NUCLEATED ERYTHROCYTES IN MATERNAL BLOOD
    BIANCHI, DW
    FLINT, AF
    PIZZIMENTI, MF
    KNOLL, JHM
    LATT, SA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (09) : 3279 - 3283
  • [10] PCR quantitation of fetal cells in maternal blood in normal and aneuploid pregnancies
    Bianchi, DW
    Williams, JM
    Sullivan, LM
    Hanson, FW
    Klinger, KW
    Shuber, AP
    [J]. AMERICAN JOURNAL OF HUMAN GENETICS, 1997, 61 (04) : 822 - 829