scholarly journals Microfluidic chip combined with magnetic-activated cell sorting technology for tumor antigen-independent sorting of circulating hepatocellular carcinoma cells

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6681 ◽  
Author(s):  
Xuebin Wang ◽  
Liying Sun ◽  
Haiming Zhang ◽  
Lin Wei ◽  
Wei Qu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Tumor Antigen ◽  
Cell Sorting ◽  
Blood Cells ◽  
Lateral Displacement ◽  
Laser Scanning Microscopy ◽  
Mesenchymal Transition ◽  
Microfluidic Structure ◽  
Magnetic Activated Cell Sorting

Purpose We aimed to generate a capture platform that integrates a deterministic lateral displacement (DLD) microfluidic structure with magnetic-activated cell sorting (MACS) technology for miniaturized, efficient, tumor antigen-independent circulating tumor cell (CTC) separation. Methods The microfluidic structure was based on the theory of DLD and was designed to remove most red blood cells and platelets. Whole Blood CD45 MicroBeads and a MACS separator were then used to remove bead-labeled white blood cells. We established HepG2 human liver cancer cells overexpressing green fluorescent protein by lentiviral transfection to simulate CTCs in blood, and these cells were then used to determine the CTC isolation efficiency of the device. The performance and clinical value of our platform were evaluated by comparison with the Abnova CytoQuest™ CR system in the separating of blood samples from 12 hepatocellular carcinoma patients undergoing liver transplantation in a clinical follow-up experiment. The isolated cells were stained and analyzed by confocal laser scanning microscopy. Results Using our integrated platform at the optimal flow rates for the specimen (60 µl/min) and buffer (100 µl/min per chip), we achieved an CTC yield of 85.1% ± 3.2%. In our follow-up of metastatic patients, CTCs that underwent epithelial–mesenchymal transition were found. These CTCs were missed by the CytoQuest™ CR bulk sorting approach, whereas our platform displayed increased sensitivity to EpCAMlow CTCs. Conclusions Our platform, which integrates microfluidic and MACS technology, is an attractive method for high-efficiency CTC isolation regardless of surface epitopes.

scholarly journals Isolation of cancer stem-like cells from hepatocellular carcinoma cell line HepG2 by methods of magnetic-activated cell sorting, spheroid culture, and anti-tumor drug-resistant selection: A primary evaluation

2020 ◽  
Vol 7 (1) ◽  
pp. 279-289
Author(s):  
Sinh Truong Nguyen ◽  
Luong Sy Nguyen ◽  
Thao Hoang Phuong Nguyen ◽  
Phuc Hong Vo ◽  
Nghia Minh Do ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Line ◽  
Cell Sorting ◽  
Carcinoma Cell ◽  
Carcinoma Cell Line ◽  
Spheroid Culture ◽  
Hepatocellular Carcinoma Cell Line ◽  
Hepatocellular Carcinoma Cell ◽  
Carcinoma Cell Line Hepg2 ◽  
Magnetic Activated Cell Sorting

Introduction: Recently reported data have suggested that only a small subset of cancer cells possess the capability to initiate malignancies. These observations were based on investigation of cells within the primary tumors displaying a distinct surface marker pattern. CD133 marker is a putative hematopoietic and neuronal stem cell marker, which is also considered to be a tumorigenic marker in brain, prostate and liver. Recent studies have shown that a small population of CD133-positive cells, indeed, exists in human hepatocellular carcinoma (HCC) cell lines and primary HCC tissues. This study was aimed at isolating the cancer stem-like cells from hepatocellular carcinoma cell line HepG2 using three different methods: magnetic-activated cell sorting (MACS), spheroid culture (SC), and anti-tumor drug (ATD) resistant selection. Methods: HepG2 hepatocellular carcinoma cells were expanded to yield enough cells that could be used to isolate cancer stem-like cells by these three methods. For MACS, cancer stem-like cells were sorted using anti-CD133 monoclonal antibody. For the second method, cancer stem-like cells were enriched by selection of anti-tumor drug resistance property. Lastly, for the third method, three-dimensional (3D) culture was used to enrich for the cancer stem-like cells. The cells obtained by the three methods were expanded to obtain an adequate number of cells for confirmation of CD133 expression. Results: The expression of CD133+ cells in the three methods was found to be different. In the MACS method, the expanded CD133+ sorted cells cultured through 2 passages only contained 0.40 % CD133+ cells. In the 3D spheroid cell culture, of the population of cells there were 38.39 % that were CD133+ cells. Lastly, in the anti-tumor drug (doxorubicin at 150 nM) resistant selection, 66.22 % were CD133+ cells. Conclusion: This study shows that isolation of HepG2 derived CD133+ population by culture with doxorubicin (150 nM) yields the highest efficiency and purity of the 3 methods studied.

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

2020 ◽  
Vol 34 (9) ◽  
Author(s):  
Dragos Nemescu ◽  
Daniela Constantinescu ◽  
Vlad Gorduza ◽  
Alexandru Carauleanu ◽  
Lavinia Caba ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Cell Sorting ◽  
Blood Cells ◽  
Maternal Blood ◽  
Nucleated Red Blood Cells ◽  
Magnetic Activated Cell Sorting

scholarly journals Twist Expression in Circulating Hepatocellular Carcinoma Cells Predicts Metastasis and Prognoses

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Liang-Chun Yin ◽  
Zhen-Chao Luo ◽  
Yan-Xin Gao ◽  
Yang Li ◽  
Qing Peng ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Epithelial Mesenchymal Transition ◽  
Critical Role ◽  
Prognostic Indicator ◽  
Tnm Staging ◽  
Mesenchymal Transition ◽  
Enrichment Technique ◽  
Tumor Number ◽  
Twist Expression

Hepatocellular carcinoma (HCC) is one of the leading malignancies worldwide. Enumeration of circulating tumor cells (CTCs) has been demonstrated to be a prognostic indicator in HCC. Twist plays a critical role in metastasis and has been proposed as a biomarker for epithelial-mesenchymal transition (EMT). However, links between the expression of Twist in CTCs and HCC clinical parameters are still unclear. This study aims to evaluate the relationship between Twist expression in CTCs and clinicohistopathological risk factors of HCC. Between June 2015 and July 2017, 80 HCC patients and 10 healthy volunteers were enrolled in this study. CTCs were isolated and analyzed by the optimized CanPatrol™ CTC-enrichment technique. Our analysis showed that Twist+ CTCs were detected in 54 of the 80 (67.5%) HCC patients. The positive ratios of Twist+ CTCs correlated with portal vein tumor thrombi, TNM staging, AFP, cirrhosis, tumor number, tumor size, and microvascular invasion. Meanwhile, the follow-up results of the 33 HCC patients who underwent hepatectomy showed that the positive ratios of Twist+ CTCs were closely correlated with the rate of metastasis or recurrence and the mortality rate. The ROC curve analyses suggested that the prognostic evaluation of Twist+ CTCs outperforms CTCs alone. Twist+ CTCs showed higher expression in Glypican-3 protein. In conclusion, Twist expression in CTCs could serve as a biomarker for evaluating HCC metastasis and prognosis.

Endogenous Heparinoids Detected by anti-Xa Activity are Present in Blood during Acute Variceal Bleeding in Cirrhosis. A Prospective Study

2014 ◽  
Vol 23 (2) ◽  
pp. 187-194 ◽  
Author(s):  
Christos Triantos ◽  
Emmanuel Louvros ◽  
Maria Kalafateli ◽  
Anne Riddell ◽  
Ulrich Thalheimer ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Variceal Bleeding ◽  
Blood Cells ◽  
Venous Pressure ◽  
White Blood Cells ◽  
International Normalized Ratio ◽  
Ulcer Bleeding ◽  
Factor X ◽  
Packed Red Blood Cells ◽  
Bacterial Peritonitis

Background & Aims: Endogenous heparinoids have been detected by thromboelastography and quantified by clotting based anti-Xa activity assays in patients with cirrhosis, but their presence in variceal bleeding has not been established yet.Methods: Clotting based anti-Xa activity was measured in A) 30 cirrhotics with variceal bleeding, B) 15 noncirrhotics with peptic ulcer bleeding, C) 10 cirrhotics without infection or bleeding, and D) 10 cirrhotics with hepatocellular carcinoma (HCC).Results: Anti-Xa activity was not detected in ulcer bleeders or in cirrhotics without infection or bleedingbut was present in seven (23%) variceal bleeders (median levels: 0.03 u/mL (0.01-0.07)) and was quantifiable for 3 days in six of seven patients. Four of seven variceal bleeders with anti-Xa activity present had HCC (p=0.023). Age, creatinine, platelet count and total infections the second day from admission were significantly correlated with the presence of measureable anti-Xa levels (p=0.014, 0.032, 0.004 and 0.019, respectively). In the HCC group, anti-Xa activity was present in three patients (30%) [median levels: 0.05 u/mL (0.01-0.06)].Conclusions: In this study, variceal bleeders and 30% of the patients with HCC had endogenous heparinoids that were detected by a clotting based anti-Xa activity assay, whereas there was no anti Xa activity present in patients with cirrhosis without infection, or bleeding or HCC, nor in those with ulcer bleeding. Thus, the anti-Xa activity is likely to be a response to bacterial infection and/or presence of HCC in cirrhosis.List of abbreviations: AFP, alpha-fetoprotein; aPTT, activated partial thromboplastin time; CP, Child-Pugh; FXa, activated factor X; GAGS, glycosaminoglycans; Hb, hemoglobin; HCC, hepatocellular carcinoma; HVPG, hepatic venous pressure gradient; INR, International normalized ratio; LMWHs, low molecular weight heparins; MELD, Model for End-stage Liver Disease; PPP, platelet-poor plasma; PRBC, packed red blood cells; PT, prothrombin time; SBP, sponataneous bacterial peritonitis; TEG, thromboelastography; WBC, white blood cells.

Sign in / Sign up

Export Citation Format

Share Document