Evaluation of the impact of density gradient centrifugation on fetal cell loss during enrichment from maternal peripheral blood

Emad, Ahmed; Drouin, Régen

doi:10.1002/pd.4387

Cited by 12 publications

(11 citation statements)

References 54 publications

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“…The Drouin group in Canada relied heavily on brute force manual microscopic scanning of all nucleated cells using primary FISH for a Y chromosome and confirmatory reverse FISH. They concluded that there were 1–3 fetal cells per cc of maternal blood and suggested that the real number may be significantly higher [Emad and Drouin, ]. Laird Jackson shared with us some unpublished data of his own from this timeframe documenting that Y positive cells detected by FISH were present in 20 out of 20 male pregnancies, further confirming that fetal cells were always or almost always present in the mother's blood.…”

Section: Fetal Cells For Prenatal Diagnosismentioning

confidence: 87%

Using fetal cells for prenatal diagnosis: History and recent progress

Beaudet

2016

American J of Med Genetics Pt C

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The potential to use fetal cells in the mother's circulation during the first or second trimester for prenatal diagnosis was described in 1968, but it has not been possible do develop a routine clinical prenatal test despite extensive commercial and academic research efforts. Early attention focused on the detection of aneuploidy, but more recent technology opens the possibility of high resolution detection of copy number abnormalities and even whole genome or exome sequencing to detect both inherited and de novo mutations. In the interim, cell-free noninvasive prenatal testing NIPT has allowed improved detection of aneuploidy, but this has led to a sharp reduction in the number of amniocentesis and chorionic villus sampling (CVS) procedures, which inevitably implies reduced detection of serious de novo deletion abnormalities. Attention has focused of both fetal nucleated red blood cells (fnRBCs) and trophoblasts. Recent progress presented at meetings, but not yet published, suggests that it will soon be possible to perform genome-wide relatively high resolution detection of deletions and duplications by recovering fetal trophoblasts during the first trimester and analyzing them by whole gene genome amplification followed by copy number analysis using arrays or next generation sequencing. © 2016 Wiley Periodicals, Inc.

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Section: Fetal Cells For Prenatal Diagnosismentioning

confidence: 87%

Using fetal cells for prenatal diagnosis: History and recent progress

Beaudet

2016

American J of Med Genetics Pt C

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“…Density gradient centrifugation is also commonly used in the enrichment of fetal cells. However, it does not provide a high enrichment, can damage cells and is associated with significant cell loss . In one instance it was reported that density gradient centrifugation resulted in the loss of 60–80% of the fetal cells and that the procedure resulted in cellular degradation, therefore, it was recommended to improve upon this, that aggressive manipulations should be eliminated and different gradients (other than Histopaque 1.119 g mL −1 ) and densities should be tested to minimize cell loss.…”

Section: Discussionmentioning

confidence: 99%

“…However, it does not provide a high enrichment, can damage cells and is associated with significant cell loss . In one instance it was reported that density gradient centrifugation resulted in the loss of 60–80% of the fetal cells and that the procedure resulted in cellular degradation, therefore, it was recommended to improve upon this, that aggressive manipulations should be eliminated and different gradients (other than Histopaque 1.119 g mL −1 ) and densities should be tested to minimize cell loss. Combining density gradient with a negative enrichment technique such as RosetteSep with antibodies such as anti‐CD45 and/or anti‐ CD36 to crosslink and cause the sedimentation of WBC can vastly decrease maternal cell contamination .…”

Section: Discussionmentioning

confidence: 99%

Isolation of Circulating Fetal Trophoblasts Using Inertial Microfluidics for Noninvasive Prenatal Testing

Winter

Hardy

Rezaei

et al. 2018

Adv Materials Technologies

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While noninvasive prenatal testing based on cell‐free fetal DNA has recently revolutionized the field of aneuploidy screening in pregnancy, it remains limited to aneuploidy and microdeletion screening, and is unable to reliably detect single gene disorders. A number of recent studies have demonstrated the potential of circulating trophoblastic cells in providing cell‐based noninvasive diagnosis with sequencing or array‐based assays. However, considering the extreme rarity of these cells in blood, efficient, high‐throughput, and clinically applicable enrichment technologies are yet to be developed. This study demonstrates for the first time the utility of inertial microfluidics for efficient isolation of trophoblastic cells from maternal peripheral blood. Under optimal operating conditions, high‐recovery yields (79%) are obtained using a trophoblastic cell‐line, which is subsequently confirmed with analysis of maternal blood. Feasibility of obtaining a diagnosis from cells isolated from a maternal sample is demonstrated in a case of confirmed fetal trisomy 21 in which six fetal cells are found in a 7 mL blood sample using fluorescence in situ hybridization. Finally, it is demonstrated that trophoblastic cells isolated using inertial microfluidics could be picked and subjected to a clinically validated sequencing assay, paving the way for further validation of this technology and larger clinical studies.

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“…This has sparked renewed interest by several groups in the isolation and analysis of intact fetal cells from maternal blood 138– 145 , which contain a pure unmixed fetal genome, with a theoretical ability for similar diagnostic accuracy as that obtained through invasive diagnostic procedures. There is strong evidence that fetal cells can be recovered and analyzed, but the approach is currently labor intensive and costly and has not yet been proven to be robustly successful and adaptable to a high-throughput, relatively low-cost diagnostic testing option.…”

Section: Concluding Remarks and Forecasts For The Future Of Prenatal mentioning

confidence: 99%

Recent advances in prenatal genetic screening and testing

Veyver

2016

F1000Res

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The introduction of new technologies has dramatically changed the current practice of prenatal screening and testing for genetic abnormalities in the fetus. Expanded carrier screening panels and non-invasive cell-free fetal DNA-based screening for aneuploidy and single-gene disorders, and more recently for subchromosomal abnormalities, have been introduced into prenatal care. More recently introduced technologies such as chromosomal microarray analysis and whole-exome sequencing can diagnose more genetic conditions on samples obtained through amniocentesis or chorionic villus sampling, including many disorders that cannot be screened for non-invasively. All of these options have benefits and limitations, and genetic counseling has become increasingly complex for providers who are responsible for guiding patients in their decisions about screening and testing before and during pregnancy.

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Evaluation of the impact of density gradient centrifugation on fetal cell loss during enrichment from maternal peripheral blood

Abstract: Data obtained raise questions about the appropriateness of the DGC step for the enrichment of rare FCs and argues for the use of the alternative nonaggressive version of the procedure presented here or prioritizing other methods of enrichments.

Cited by 12 publications

References 54 publications

Using fetal cells for prenatal diagnosis: History and recent progress

Using fetal cells for prenatal diagnosis: History and recent progress

Isolation of Circulating Fetal Trophoblasts Using Inertial Microfluidics for Noninvasive Prenatal Testing

Recent advances in prenatal genetic screening and testing

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