A Silicon-based Coral-like Nanostructured Microfluidics to Isolate Rare Cells in Human Circulation: Validation by SK-BR-3 Cancer Cell Line and Its Utility in Circulating Fetal Nucleated Red Blood Cells

Ma, Gwo‐Chin; Lin, Wen‐Hsiang; Huang, Chihpin; Chang, Ting‐Yu; Liu, Jiayun; Yang, Yajun; Lee, Mei‐Hui; Wu, Wan‐Ju; Chang, Ya‐Hsuan; Chen, Ming

doi:10.3390/mi10020132

Cited by 20 publications

(29 citation statements)

References 44 publications

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“…In recent years, many approaches based on microfluidics chips have been developed to capture circulating fetal cells. [13][14][15][16][17] Despite considerable progress, reproducibility, and reliability of isolation and detection of fetal cells from maternal blood remain poor. 18 That was attributed to the rarity and variability of fetal cells among pregnancies.…”

Section: Introductionmentioning

confidence: 99%

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Comparison between paramagnetic and CD71 magnetic activated cell sorting of fetal nucleated red blood cells from the maternal blood

Nemescu

Constantinescu

Gorduza

et al. 2020

Clinical Laboratory Analysis

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Background Fetal nucleated red blood cells (NRBC) from maternal circulation are rare events but can be enriched and used to evaluate the genetics of the fetus. We compared two simplified selection methods of the fetal cells from the maternal blood. Methods We isolated fetal cells from maternal blood through double‐density gradient centrifugation followed either by magnetic cell selection, based on the paramagnetic proprieties of the NRBC hemoglobin, converted to methemoglobin, or by a positive magnetic‐activated cell sorting (MACS) enrichment, using anti‐CD71 monoclonal antibodies. Finally, the cells were identified through fluorescence in situ hybridization (FISH) with specific chromosome X and Y probes. Results We processed 10 mL of peripheral blood samples from 27 pregnant women with singleton normal male fetuses. Hemoglobin‐based enrichment isolated significantly more NRBCs: 29.7 × 104 cells than anti‐CD71 MACS: 10.1 × 104 cells (P < .001). The FISH analysis found at least one XY cell in 81.5% and 61.5% of cases, respectively, for paramagnetic and anti‐CD71 selection. Also, the average number of XY cells identified through paramagnetic selection was 5.09 ± 2.5, significantly higher than those observed through CD71 sorting: 3.38 ± 1.7 cells (average ± SE) (P = .03). Conclusion The combination of density gradient centrifugation with paramagnetic selection has the advantage of simplicity and achieves a minimal manipulation and treatment of cells. It yields an increased number of NRBCs and FISH confirmed fetal cells, compared to the anti‐CD71 sorting.

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Section: Introductionmentioning

confidence: 99%

“…However, the procedures are complex that limits extensive studies. In recent years, many approaches based on microfluidics chips have been developed to capture circulating fetal cells 13‐17 …”

Section: Introductionmentioning

confidence: 99%

Comparison between paramagnetic and CD71 magnetic activated cell sorting of fetal nucleated red blood cells from the maternal blood

Nemescu

Constantinescu

Gorduza

et al. 2020

Clinical Laboratory Analysis

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“…However, the detection of fNRBCs is challenging due to their extremely low concentration against a background predominance of maternal cells (<6 cells per mL; with 109 maternal cells) 16,17 . Several fNRBC enrichment methods based on different principles have been reported, such as density gradient centrifugation (DGC) 13,18 , fluorescence-activated cell sorting (FACS) 19 , and magnetic-activated cell sorting (MACS) 20 , dielectrophoresis, and microfluidics based technologies 21,22 . Nevertheless, long-term preservation of samples and rapid identification of target cells (fNRBCs) still present considerable challenges.…”

Section: Introductionmentioning

confidence: 99%

Deep learning-based adaptive detection of fetal nucleated red blood cells

Sun

Wang

Zhao

et al. 2020

Preprint

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15Aim: this study, we established an artificial intelligence system for rapid 16 identification of fetal nucleated red blood cells (fNRBCs). 17Method: Density gradient centrifugation and magnetic-activated cell sorting were 18 used for the separation of fNRBCs from umbilical cord blood. The cell block 19 technique was used for fixation. We proposed a novel preprocessing method based on 20 imaging characteristics of fNRBCs for region of interest (ROI) extraction, which 21 automatically segmented individual cells in peripheral blood cell smears. The 22 discriminant information from ROIs was encoded into a feature vector and 23 pathological diagnosis were provided by the prediction network. 24Results: Four umbilical cord blood samples were collected and validated based on a 25 large dataset containing 260 samples. Finally, the dataset was classified into 3,720 and 26 1,040 slides for training and testing, respectively. In the test set, classifier obtained 27 98.5% accuracy and 96.5% sensitivity. 28Conclusion: Therefore, this study offers an effective and accurate method for 29 fNRBCs preservation and identification. 30 Keywords: fetal nucleated red blood cells, cell-block, deep learning, non-invasive 31 prenatal diagnosis 32 Introduction 33The clinical application of fNRBCs during pregnancy could be classified into two 34 main categories 1,2 . One is the prognosis of possible diseases in pregnant women by 35 counting fNRBCs in umbilical cord blood. Chronic tissue hypoxia results in increased 36 levels of erythropoietin, which, in turn, leads to stimulation of erythropoiesis and 37 increased numbers of circulating nucleated red blood cells (NRBCs) 1,3-5 . Increased 38 umbilical cord levels of erythropoietin have been reported in pregnancies complicated 39 by intrauterine growth restriction, maternal hypertension, preeclampsia, maternal 40 smoking, Rh isoimmunization, and maternal diabetes 5-8 . As expected, each of these 41 conditions has been associated with increased NRBCs in the newborn 9 . The other 42 objective is to screen and extract fNRBCs from maternal peripheral blood for 43 non-invasive prenatal diagnosis (NIPD) 10-12 . The choice of fNRBCs as ideal target 44 cells is based on the following parameters 13,14 : (1) presence of intact nuclei containing 45 the complete fetal genome in fNRBCs, which is a prerequisite for prenatal analysis; (2) 46 limited life span of fNRBCs in the maternal circulation, which can be differentiated 47 55 cells) 16,17 . Several fNRBC enrichment methods based on different principles have 56 been reported, such as density gradient centrifugation (DGC) 13,18 , 57 fluorescence-activated cell sorting (FACS) 19 , and magnetic-activated cell sorting 58 (MACS) 20 , dielectrophoresis, and microfluidics based technologies 21,22 . Nevertheless, 59 long-term preservation of samples and rapid identification of target cells (fNRBCs) 60 still present considerable challenges. 61 Since the identification of fNRBCs in large number of cell block slices represent a 62 huge manual burden on p...

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“…This volume highlights a diverse collection of research on single cell manipulation, diagnostics, cell migration, cell flow cytometry, to name a few. I am also happy to see that some papers included automated systems to operate the devices [3,4]. Automation is needed if we want to have a more intensive use of microfluidic based platforms and reproducibility.…”

mentioning

confidence: 99%

“…As it turns out, circulating fetal cells (CFCs) are present in the maternal blood. Ma et al [4] designed a chip and an automated system to isolate this rare cell from the maternal blood.…”

mentioning

confidence: 99%