Identification of fetal nucleated red cells in co‐cultures from fetal and adult peripheral blood: differential effects of serum on fetal and adult erythropoiesis

Böhmer, Ralph M.; Zhen, DongKai; Bianchi, Diana W.

doi:10.1002/(sici)1097-0223(199907)19:7<628::aid-pd601>3.0.co;2-#

Cited by 25 publications

(4 citation statements)

References 24 publications

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“…Cultures were established in standard 6-well plates, 3 ml per well, at a maximum density of 0.2r10 6 mononuclear cells/ml. The standard medium was a mixture of O Iscoves MDM and M RPMI1640, containing methylcellulose (0.9%), recombinant human erythropoietin (EPO, 1 U/ml), recombinant human stem cell factor (SCF, 20 ng/ml), insulin (3 mg/ml), iron-saturated transferrin (70 mg/ml), mercaptoethanol (0.7 mM) and 1% charcoal-stripped human cord serum, prepared as described previously (Bohmer et al, 1999). Recombinant human interleukin-3 (IL3) was added at 20 ng/ml.…”

Section: Cell Culturementioning

confidence: 99%

“…Optimizing fetal cell purity by manipulating culture conditions depends on agents that affect fetal and adult F+Ax cells differentially. We have shown previously that the growth of adult F+Ax cells can be reduced selectively by using charcoal-treated human cord serum at low concentrations (Bohmer et al, 1999). However, a small amount of adult F+Ax cells remained after optimizing the serum content.…”

Section: Introductionmentioning

confidence: 99%

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Differential effects of interleukin‐3 on fetal and adult erythroid cells in culture: implications for the isolation of fetal cells from maternal blood

Böhmer¹,

Johnson²,

Bianchi³

2000

Prenat. Diagn.

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Fetal clonogenic erythroid cells may be present in maternal blood and serve as a source of fetal DNA for prenatal genetic diagnosis. Proliferating nucleated red cells in cultures from first and second trimester fetal blood contain only fetal haemoglobin (HbF; F+A- cells), whereas nucleated red cells from adult blood contain also adult haemoglobin (HbA; F+A+ or F-A+ cells). Thus, fetal red cells can be identified and flow sorted. However, a few adult cells are also F+A-, which reduces the purity of fetal cell isolation. Culture media optimized for erythropoiesis contain interleukin-3 (IL3). We show here that IL3 strongly stimulates the growth of F+ cells (both F+A- and F+A+) in cultures from adult blood but has only a comparatively small effect on F+A- cells in cultures from fetal blood. This difference is maintained in the unified conditions of co-cultures of adult and fetal cells, so that the purity of fetal cell sorts can be increased by omitting IL3 from the culture medium. We further show that IL3 accelerates the exhaustion of the long-term division potential of adult cells, allowing fetal secondary colonies to be identified by their size following a two-stage culture scheme. Thus, the choice to omit or include IL3 in the growth medium of maternal blood cultures should depend on whether fetal nucleated red cells are to be isolated by flow sorting after one week, or by picking secondary colonies after a later secondary culture stage.

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Section: Cell Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Differential effects of interleukin‐3 on fetal and adult erythroid cells in culture: implications for the isolation of fetal cells from maternal blood

Böhmer¹,

Johnson²,

Bianchi³

2000

Prenat. Diagn.

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“…In addition to the difficulty of selecting fetal cells of hematopoietic lineage, their in vitro expansion has also yielded disappointing results (Chen et al, 1998). Despite the development of seemingly optimal fetal cell culture conditions, and identification of optimal in vitro cytokine combinations that favor the proliferation of fetal over adult progenitor cells, fetal cells from maternal blood samples thus far have not been consistently expanded (Bohmer et al, 1999(Bohmer et al, , 2000(Bohmer et al, , 2002Manotaya et al, 2002;Zimmermann et al, 2002;. Fetal mesenchymal stem cells in maternal circulation have also been investigated as possible targets, but their rarity makes them unlikely candidate cells for prenatal diagnosis (Campagnoli et al, 2001;O'Donoghue et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Culture of endothelial cells isolated from maternal blood using anti‐CD105 and CD133

2004

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“…Moreover, fetal status may also affect the feto-maternal transfusion. Most investigators start the NRBC enrichment with a single, double or triple density gradient, ranging from 1.077 to 1.119 g/ml (13)(14)(15)(16). Different enrichment techniques have been proposed to separate fetal from maternal cells (3,8,9).…”

Section: Introductionmentioning

confidence: 99%

Comparison of Different CD71 Monoclonal Antibodies for Enrichment of Fetal Cells from Maternal Blood

Prieto

Cándenas²,

Ladenson³

et al. 2002

Clinical Chemistry and Laboratory Medicine

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Different approaches have been proposed for the enrichment of fetal nucleated red blood cells (NRBC) from maternal blood as an alternative way to obtain fetal tissue for non-invasive prenatal diagnosis. The main purpose of this study was to compare two of our monoclonal antibodies (2E11.3 and 2B7.4 mAbs) with the most widely used commercial anti-CD71 mAb, in terms of their ability to isolate NRBC from maternal blood by magnetic activated cell sorting (MACS). Peripheral blood samples were obtained from 60 pregnant women at a mean gestational age of 16 weeks (range: 10-19 weeks). The number of NRBC isolated by our antibodies (median: 68, range: 0-2102) was significantly higher than that obtained by the commercial antibody (median: 38, range: 0-2165) in the same samples. However, in the final preparations, contamination by maternal nucleated blood cells was lower when the commercial antibody was used. Since fetal NRBC are rare in maternal blood, the improved NRBC recovery achieved by our non-commercial antibodies should facilitate the non-invasive detection of fetal aneuploidies in maternal blood.

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Identification of fetal nucleated red cells in co‐cultures from fetal and adult peripheral blood: differential effects of serum on fetal and adult erythropoiesis

Cited by 25 publications

References 24 publications

Differential effects of interleukin‐3 on fetal and adult erythroid cells in culture: implications for the isolation of fetal cells from maternal blood

Differential effects of interleukin‐3 on fetal and adult erythroid cells in culture: implications for the isolation of fetal cells from maternal blood

Culture of endothelial cells isolated from maternal blood using anti‐CD105 and CD133

Comparison of Different CD71 Monoclonal Antibodies for Enrichment of Fetal Cells from Maternal Blood

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