Flow cytometric methods for prenatal and neonatal diagnosis

Curtis, Michèle G.; Walker, Brooke; Denny, Thomas N.

doi:10.1016/j.jim.2010.09.039

Cited by 10 publications

(4 citation statements)

References 63 publications

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“…Moreover, FCM-based cell sorting takes advantage of the same reagents and expertise as FCM analysis and is already being applied for diagnostic purposes. 50 , 51 …”

Section: Discussionmentioning

confidence: 99%

Identification and surveillance of rare relapse-initiating stem cells during complete remission after transplantation

Dimitriou,

Mortera-Blanco,

Tobiasson

et al. 2024

Blood

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Relapse following complete remission (CR) remains the main cause of mortality after allogeneic stem cell transplantation for hematological malignancies and therefore improved biomarkers for early prediction of relapse remains a critical goal towards development and assessment of preemptive relapse treatment. Since the significance of cancer stem cells as a source of relapses remains unclear, we investigated whether mutational screening for persistence of rare cancer stem cells would enhance measurable residual disease (MRD) and early relapse-prediction post-transplantation. In a retrospective study of relapse patients and continuous-CR patients with myelodysplastic syndromes and related myeloid malignancies, combined flow cytometric cell sorting and mutational screening for persistence of rare relapse-initiating stem cells was performed in bone marrow at multiple CR time points post-transplantation. In 25 CR samples from 15 patients that later relapsed, only 9 samples were MRD-positive in mononuclear cells (MNCs) whereas flowcytometric sorted hematopoietic stem and progenitor cells (HSPCs) were MRD-positive in all samples, and always with a higher variant allele frequency than in MNCs (mean 97-fold). MRD-positivity in HSPCs preceded MNCs in multiple sequential samples, in some cases preceding relapse by more than 2 years. In distinction, in 13 patients in long-term continuous-CR, HSPCs remained MRD-negative. Enhanced MRD-sensitivity was also observed in total CD34+ cells, but HSPCs were always more clonally involved (mean 8-fold).In conclusion, identification of relapse-initiating cancer stem cells and mutational MRD-screening for their persistence consistently enhances MRD-sensitivity and earlier prediction of relapse after allogeneic stem cell transplantation.

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“…Moreover, FCM-based cell sorting takes advantage of the same reagents and expertise as FCM analysis and is already being applied for diagnostic purposes. 50 , 51 …”

Section: Discussionmentioning

confidence: 99%

Identification and surveillance of rare relapse-initiating stem cells during complete remission after transplantation

Dimitriou,

Mortera-Blanco,

Tobiasson

et al. 2024

Blood

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“…Flow cytometric procedure enumerates CD3+ cells (T lymphocytes), CD3+CD4+ cells (T helper cells), CD3+CD8+ cells (T cytotoxic cells), CD19+ or CD20+ cells (B lymphocytes), and CD3-CD16+/56+ cells (NK cells). This test will discover most infants with SCID or complete DiGeorge syndrome and may give guidance as to the character of the T-cell-related defect [ 19 , 20 ]. If a T-cell defect is thought, the preliminary test for T-cell function is a lymphocyte proliferation assay.…”

Section: Initial Laboratory Evaluation For Primary Immunodeficiencies Of Newbornmentioning

confidence: 99%

“…Documentation of a mutated Bruton’s tyrosine kinase (BTK) gene confirms the most commonly seen X-linked agammaglobulinemia. If an infant is a girl or the BTK gene is normal, autosomal recessive (AR) agammaglobulinemia should be considered [ 19 , 27 ].…”

Section: Most Common Types Of Pids In Newbornmentioning

confidence: 99%

Primary Immunodeficiency Diseases in the Newborn

Özdemir¹

2020

North Clin Istanbul

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The normal neonate’s immune system is anatomically completed but antigenically inexperienced and shows somewhat decreased role of a number of immunological pathways. Aside from anatomic characteristics (e.g., thin skin and mucosal barriers) of newborn, weakened pro-inflammatory and T-helper cell type 1 cytokine release and lessened cell-mediated immunity predispose the neonate more susceptible to all types of infections. Furthermore, many types of primary immunodeficiency diseases (PIDs) that present in neonatal period are potentially life threatening. However, most of the newborns stand this period without sickness due to complete innate immunity with other adaptive immune system mechanisms and transferred maternal immunoglobulin G. Besides unique immunity of the preterm and normal newborns; risk factors, clinical features, and laboratory evaluation of most common PIDs in newborn are told in this article. The range of PIDs is growing, and the diagnosis and management of these disorders continues to increase in complexity. The most common PID types of the newborn including antibody deficiencies, cellular/combined immunodeficiencies, phagocytic diseases, complement deficiencies, and innate immune system and other disorders are briefly mentioned here as well.

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“…Considering that fetal nucleated erythroblasts are rare in the maternal blood, it is essential to develop a reliable and stable approach to isolate sufficient numbers from the maternal blood circulation for noninvasive prenatal diagnosis. Even though several methods such as fluorescence-activated cell sorting (FACS) [12], magnetic-activated cell sorting (MACS) [12], charge-flow separation [13] and lectins [14] have been adopted for the isolation of fetal cells, the reliability and efficiency of these methods remain questionable, limiting their clinical application [8,9,14,15,16,17,18]. To overcome the scarcity of fetal nucleated erythroblasts in the maternal blood, some researchers have attempted to amplify the cell colony by culturing them in vitro [19,20,21,22,23]; however, the feasibility of this culturing process for a clinical operation remains elusive, partly due to the limitation of the technique employed in the experiments [24,25,26].…”

Section: Introductionmentioning

confidence: 99%

A Comparison of in vitro Culture of Fetal Nucleated Erythroblasts from Fetal Chorionic Villi and Maternal Peripheral Blood for Noninvasive Prenatal Diagnosis

Zheng¹,

Tong²,

Wu³

et al. 2012

Fetal Diagn Ther

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Objective: We tested the feasibility of the in vitro culture of fetal nucleated erythroblasts from maternal blood for noninvasive prenatal screening as a substitute for culturing fetal nucleated erythroblasts from fetal villi. Method: Nucleated blood cells separated via Percoll from 52 samples of fetal villi and maternal peripheral blood were cultured with or without magnetic-activated cell sorting glycophorin A (MACS-GPA+), and detected by an anti-hemoglobin-epsilon (FITC) antibody. Gender of the epsilon-positive cells were identified by FISH and further confirmed by PCR of the villi karyotype. Developmental stages of nucleated erythroblasts from villi and blood with MACS-GPA+ were analyzed by Wright-Giemsa staining. Results: In the maternal blood, epsilon-positive cells were found in 4 and 24 cultured samples with and without MACS-GPA+, respectively. Also, Y-signals were visualized in 3 out of 4 and in 15 out of 24 cases in the epsilon-positive cells. Although epsilon-positive cells were found in all villus samples irrespective of MACS-GPA+ sorting, Y-signals were visualized in 31 out of 52 cases. Proerythroblasts and basophilic erythroblasts occupied 7 and 1% in fetal and maternal samples (with MACS-GPA+), respectively. Conclusions: The in vitro culturing of fetal nucleated erythroblasts from maternal blood is not feasible with the current techniques for prenatal diagnosis, because the fetal nucleated erythroblast is not well developed in vitro. This may be attributed to the low proportion of these erythroblasts at an early stage in the fetal circulation and the low permeability of these cells to the maternal blood.

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Flow cytometric methods for prenatal and neonatal diagnosis

Cited by 10 publications

References 63 publications

Identification and surveillance of rare relapse-initiating stem cells during complete remission after transplantation

Identification and surveillance of rare relapse-initiating stem cells during complete remission after transplantation

Primary Immunodeficiency Diseases in the Newborn

A Comparison of in vitro Culture of Fetal Nucleated Erythroblasts from Fetal Chorionic Villi and Maternal Peripheral Blood for Noninvasive Prenatal Diagnosis

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