scholarly journals Biocompatible and label-free separation of cancer cells from cell culture lines from white blood cells in ferrofluids

Lab on a Chip ◽  
2017 ◽  
Vol 17 (13) ◽  
pp. 2243-2255 ◽  
Author(s):  
Wujun Zhao ◽  
Rui Cheng ◽  
So Hyun Lim ◽  
Joshua R. Miller ◽  
Weizhong Zhang ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Blood Cells ◽  
White Blood Cells ◽  
Label Free ◽  
Low Concentration

A biocompatible and label-free method for separation of low-concentration cancer cells from cell lines from white blood cells is developed.

scholarly journals Discrimination between Breast Cancer Cells and White Blood Cells by Non-Invasive Measurements: Implications for a Novel In Vitro-Based Circulating Tumor Cell Model Using Digital Holographic Cytometry

2020 ◽  
Vol 10 (14) ◽  
pp. 4854
Author(s):  
Zahra El-Schich ◽  
Birgit Janicke ◽  
Kersti Alm ◽  
Nishtman Dizeyi ◽  
Jenny L. Persson ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Cell ◽  
Cancer Cells ◽  
Cell Lines ◽  
Blood Cells ◽  
Breast Cancer Cells ◽  
Cell Model ◽  
White Blood Cells ◽  
Circulating Tumor Cell

Breast cancer is the second most common cancer worldwide. Metastasis is the main reason for death in breast cancer, and today, there is a lack of methods to detect and isolate circulating tumor cells (CTCs), mainly due to their heterogeneity and rarity. There are some systems that are designed to detect rare epithelial cancer cells in whole blood based on the most common marker used today, the epithelial cell adhesion molecule (EpCAM). It has been shown that aggressive breast cancer metastases are of non-epithelial origin and are therefore not always detected using EpCAM as a marker. In the present study, we used an in vitro-based circulating tumor cell model comprising a collection of six breast cancer cell lines and white blood cell lines. We used digital holographic cytometry (DHC) to characterize and distinguish between the different cell types by area, volume and thickness. Here, we present significant differences in cell size-related parameters observed when comparing white blood cells and breast cancer cells by using DHC. In conclusion, DHC can be a powerful diagnostic tool for the characterization of CTCs in the blood.

scholarly journals In Vitro Spectroscopy-Based Profiling of Urothelial Carcinoma: A Fourier Transform Infrared and Raman Imaging Study

Cancers ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 123
Author(s):  
Monika Kujdowicz ◽  
Wojciech Placha ◽  
Brygida Mech ◽  
Karolina Chrabaszcz ◽  
Krzysztof Okoń ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Fourier Transform ◽  
Cancer Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Malignant Neoplasm ◽  
Fourier Transform Infrared ◽  
Diagnostic Tools ◽  
Label Free ◽  
Bladder Cancer Cells

Markers of bladder cancer cells remain elusive, which is a major cause of the low recognition of this malignant neoplasm and its recurrence. This implies an urgent need for additional diagnostic tools which are based on the identification of the chemism of bladder cancer. In this study, we employed label-free techniques of molecular imaging—Fourier Transform Infrared and Raman spectroscopic imaging—to investigate bladder cancer cell lines of various invasiveness (T24a, T24p, HT-1376, and J82). The urothelial HCV-29 cell line was the healthy control. Specific biomolecules discriminated spatial distribution of the nucleus and cytoplasm and indicated the presence of lipid bodies and graininess in some cell lines. The most prominent discriminators are the total content of lipids and sugar moieties as well as the presence of glycogen and other carbohydrates, un/saturated lipids, cytochromes, and a level of S-S bridges in proteins. The combination of the obtained hyperspectral database and chemometric methods showed a clear differentiation of each cell line at the level of the nuclei and cytoplasm and pointed out spectral signals which differentiated bladder cancer cells. Registered spectral markers correlated with biochemical composition changes can be associated with pathogenesis and potentially used for the diagnosis of bladder cancer and response to experimental therapies.

scholarly journals Detection of Blood Cancer Cells Using Microscopic Images

2021 ◽  
Vol 9 (8) ◽  
pp. 995-1003
Author(s):  
Vidyashree M S
Keyword(s):  
Machine Learning ◽  
Support Vector Machine ◽  
Cancer Cells ◽  
Blood Cells ◽  
White Blood Cells ◽  
Machine Learning Algorithms ◽  
Support Vector ◽  
Support Vector Machine Algorithm ◽  
Blood Cancer ◽  
Acute Myeloid

Abstract: Blood Cancer cells forming a tissue is called lymphoma. Thus, disease decreases the cells to fight against the infection or cancer blood cells. Blood cancer is also categorized in too many types. The two main categories of blood cancer are Acute Lymphocytic Lymphoma and Acute Myeloid Lymphoma. In this project proposes a approach that robotic detects and segments the nucleolus from white blood cells in the microscopic Blood images. Here in this project, we have used the two Machine learning algorithms that are k-means algorithm, Support vector machine algorithm. K-mean algorithm is use for segmentation and clustering. Support vector machine algorithm is used for classification. Keywords: k-means, Support vector machine, Lymphoma, Acute Lymphocytic Lymphoma, Machine Learning

Segmentation of Leukemia Cells Using Clustering

2019 ◽  
Vol 10 (2) ◽  
pp. 39-48
Author(s):  
Eman Mostafa ◽  
Heba A. Tag El-Dien
Keyword(s):  
Bone Marrow ◽  
Cancer Cells ◽  
Blood Cells ◽  
White Blood Cells ◽  
Leukemia Cells ◽  
Microscopic Image ◽  
Processing Stage ◽  
Clustering Techniques ◽  
Fuzzy C Means

Leukemia is a blood cancer which is defined as an irregular augment of undeveloped white blood cells called “blasts.” It develops in the bone marrow, which is responsible for blood cell generation including leukocytes and white blood cells. The early diagnosis of leukemia greatly helps in the treatment. Accordingly, researchers are interested in developing advanced and accurate automated techniques for localizing such abnormal blood cells. Subsequently, image segmentation becomes an important image processing stage for successful feature extraction and classification of leukemia in further stages. It aims to separate cancer cells by segmenting the microscopic image into background and cancer cells that are known as the region of interested (ROI). In this article, the cancer blood cells were segmented using two separated clustering techniques, namely the K-means and Fuzzy-c-means techniques. Then, the results of these techniques were compared to in terms of different segmentation metrics, such as the Dice, Jac, specificity, sensitivity, and accuracy. The results proved that the k-means provided better performance in leukemia blood cells segmentation as it achieved an accuracy of 99.8% compared to 99.6% with the fuzzy c-means.

scholarly journals Secretomic Analysis Identifies Alpha-1 Antitrypsin (A1AT) as a Required Protein in Cancer Cell Migration, Invasion, and Pericellular Fibronectin Assembly for Facilitating Lung Colonization of Lung Adenocarcinoma Cells

2012 ◽  
Vol 11 (11) ◽  
pp. 1320-1339 ◽  
Author(s):  
Ying-Hua Chang ◽  
Shu-Hui Lee ◽  
I-Chuang Liao ◽  
Shin-Huei Huang ◽  
Hung-Chi Cheng ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Cancer Cells ◽  
Cell Lines ◽  
Metastatic Potential ◽  
Cell Surface Receptor ◽  
Migration And Invasion ◽  
Secretory Proteins ◽  
Label Free ◽  
Altered Expression ◽  
Lung Colonization

Metastasis is a major obstacle that must be overcome for the successful treatment of lung cancer. Proteins secreted by cancer cells may facilitate the progression of metastasis, particularly within the phases of migration and invasion. To discover metastasis-promoting secretory proteins within cancer cells, we used the label-free quantitative proteomics approach and compared the secretomes from the lung adenocarcinoma cell lines CL1-0 and CL1-5, which exhibit low and high metastatic properties, respectively. By employing quantitative analyses, we identified 660 proteins, 68 of which were considered to be expressed at different levels between the two cell lines. High levels of A1AT were secreted by CL1-5, and the roles of A1AT in the influence of lung adenocarcinoma metastasis were investigated. Molecular and pathological confirmation demonstrated that altered expression of A1AT correlates with the metastatic potential of lung adenocarcinoma. The migration and invasion properties of CL1-5 cells were significantly diminished by reducing the expression and secretion of their A1AT proteins. Conversely, the migration and invasion properties of CL1-0 cells were significantly increased through the overexpression and secretion of A1AT proteins. Furthermore, the assembly levels of the metastasis-promoting pericellular fibronectin (FN1), which facilitates colonization of lung capillary endothelia by adhering to the cell surface receptor dipeptidyl peptidase IV (DPP IV), were higher on the surfaces of suspended CL1-5 cells than on those of the CL1-0 cells. This discovery reflects previous findings in breast cancer. In line with this finding, FN1 assembly and the lung colonization of suspended CL1-5 cells were inhibited when endogenous A1AT protein was knocked down using siRNA. The major thrust of this study is to demonstrate the effects of coupling the label-free proteomics strategy with the secretomes of cancer cells that differentially exhibit invasive and metastatic properties. This provides a new opportunity for the effective identification of metastasis-associated proteins that are secreted by cancer cells and promote experimental metastasis.

scholarly journals Microfluidic Separation of Blood Cells Based on the Negative Dielectrophoresis Operated by Three Dimensional Microband Electrodes

Micromachines ◽  
2020 ◽  
Vol 11 (9) ◽  
pp. 833
Author(s):  
Tomoyuki Yasukawa ◽  
Junko Yamada ◽  
Hitoshi Shiku ◽  
Tomokazu Matsue ◽  
Masato Suzuki
Keyword(s):  
Electric Field ◽  
Blood Cells ◽  
White Blood Cells ◽  
Three Dimensional ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Acute Monocytic Leukemia ◽  
Label Free ◽  
Monocytic Leukemia ◽  
Insulating Layer

A microfluidic device is presented for the continuous separation of red blood cells (RBCs) and white blood cells (WBCs) in a label-free manner based on negative dielectrophoresis (n-DEP). An alteration of the electric field, generated by pairs of slanted electrodes (separators) that is fabricated by covering parts of single slanted electrodes with an insulating layer is used to separate cells by their sizes. The repulsive force of n-DEP formed by slanted electrodes prepared on both the top and bottom substrates led to the deflection of the cell flow in lateral directions. The presence of gaps covered with an insulating layer for the electric field on the electrodes allows the passing of RBCs through gaps, while relatively large WBCs (cultured cultured human acute monocytic leukemia cell line (THP-1 cells)) flowed along the slanted separator without passing through the gaps and arrived at an edge in the channel. The passage efficiency for RBCs through the gaps and the arrival efficiency for THP-1 cells to the upper edge in the channel were estimated and found to be 91% and 93%, respectively.

scholarly journals Human Erythroblasts with c-Kit Activating Mutations Have Reduced Cell Culture Costs and Remain Capable of Terminal Maturation and Enucleation

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2315-2315
Author(s):  
Tyler A Couch ◽  
Zachary C. Murphy ◽  
Michael Getman ◽  
Ryo Kurita ◽  
Yukio Nakamura ◽  
...  
Keyword(s):  
Cell Culture ◽  
Red Blood Cells ◽  
Cell Lines ◽  
Blood Cells ◽  
In Vitro Production ◽  
Activating Mutations ◽  
Expansion Medium ◽  
The Cost ◽  
Vitro Production

Abstract There is a constant need for red blood cells for transfusion therapy in the treatment of anemias and acute injury. As all blood products for transfusion come from donors, there are concerns over shortages and safety. Furthermore, many patients with transfusion-dependent anemias risk alloiumminization. The in vitro production of red blood cells would address these problems, especially as they can be genetically engineered to prevent alloimmunization. Numerous erythroid culture systems now exist for the in vitro production of red blood cells. Hematopoietic stem and progenitor cells (HSPCs) obtained from umbilical cord or peripheral blood can be differentiated into erythrocytes, however, they are limited in expansion. While umbilical cord HSPCs have greater expandability than peripheral blood, the resulting erythrocytes contain fetal globins. Pluripotent stem cells can also be used as a starting source, however only a small percentage of the cells can be differentiated into erythroblasts which also suffer from low enucleation rates. Presently, the cost of in vitro production of a unit of red cells is greater than an order of magnitude higher than obtaining it from a donor largely due to the medium and cytokine costs (Timmins & Nielsen, Trends Biotechnol, 2009). A relatively new approach of immortalizing early erythroblasts allowing unlimited expansion as well as terminal maturation and enucleation shows great therapeutic promise (Kurita et al., PLoS One, 2013; Huang et al., Mol Ther, 2014; Trakarnsanga et al., Nat Commun, 2017). However, these immortalized erythroblasts are still reliant on two costly cytokines: stem cell factor (SCF) and erythropoietin (Epo). Mutations in exon 17 of the receptor tyrosine kinase gene KIT are frequently seen in acute myeloid leukemias, gastrointestinal stromal tumors, and mast cells leading to mastocytosis. These mutations cause the c-Kit protein to spontaneously activate and transduce signal in the absence of SCF (Kit-ligand). To generate an SCF-independent HUDEP-2 cell line (Kurita et al., PLoS One, 2013), we used CRISPR/Cas9 to introduce missense and frameshifting mutations within the vicinity of Asp816 in exon 17 of the KIT gene. The resulting monoclonal cell lines were selected for by removing SCF from the expansion medium and were subsequently named KIT-CAT (KIT with Constitutively Activating Transformation). To better understand what KIT mutations allowed or impaired terminal maturation, monoclonal cell lines were genotyped by Sanger sequencing. Three cell lines with unique genotypes were chosen for further analysis. All three KIT-CAT lines had a shorter doubling time compared to HUDEP-2 cells (16.7 vs 18.9 hrs, p=0.020) and were no longer dependent on SCF or Epo. However, two of the three KIT-CAT lines showed more robust proliferation with Epo in the expansion medium. The addition of SCF to the medium caused no increase in c-Kit activation by Western blotting for phosphorylation at Tyr703. Furthermore, the low molecular weight and immature form of c-Kit is also phosphorylated in KIT-CAT cells, but not HUDEP-2 cells, indicating c-Kit activation occurs before trafficking to the cell membrane where SCF would bind (Tabone-Eglinger et al., Clin Cancer Res, 2008). Key features of erythroblast maturation are the decrease in cell and nuclear size which can be measured using imaging flow cytometry (McGrath et al., Methods, 2017). While in expansion phase, all 3 cell lines were larger in cell and nuclear area compared to the parental HUDEP-2 line. By day 6 of maturation, all three cell lines had statistically significant decreases in cell and nuclear size indicating maturation. By day 13 of culture, Wright-Giemsa staining showed that the majority of the cells were orthochromatic erythroblasts or enucleate reticulocytes. Reducing cell culture costs is needed for in vitro manufacturing of red blood cells to be economically feasible. These results show that a c-Kit activating mutations in human erythroblasts removes the cost of SCF and reduces the cost of Epo while still allowing for terminal maturation and enucleation. Disclosures No relevant conflicts of interest to declare.

scholarly journals The landscape of metabolic pathway dependencies in cancer cell lines

2021 ◽  
Vol 17 (4) ◽  
pp. e1008942
Author(s):  
James H. Joly ◽  
Brandon T. L. Chew ◽  
Nicholas A. Graham
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Metabolic Pathway ◽  
Culture Medium ◽  
Cancer Cell Lines ◽  
Metabolic Reprogramming ◽  
Cell Culture Medium ◽  
Loss Of Function

The metabolic reprogramming of cancer cells creates metabolic vulnerabilities that can be therapeutically targeted. However, our understanding of metabolic dependencies and the pathway crosstalk that creates these vulnerabilities in cancer cells remains incomplete. Here, by integrating gene expression data with genetic loss-of-function and pharmacological screening data from hundreds of cancer cell lines, we identified metabolic vulnerabilities at the level of pathways rather than individual genes. This approach revealed that metabolic pathway dependencies are highly context-specific such that cancer cells are vulnerable to inhibition of one metabolic pathway only when activity of another metabolic pathway is altered. Notably, we also found that the no single metabolic pathway was universally essential, suggesting that cancer cells are not invariably dependent on any metabolic pathway. In addition, we confirmed that cell culture medium is a major confounding factor for the analysis of metabolic pathway vulnerabilities. Nevertheless, we found robust associations between metabolic pathway activity and sensitivity to clinically approved drugs that were independent of cell culture medium. Lastly, we used parallel integration of pharmacological and genetic dependency data to confidently identify metabolic pathway vulnerabilities. Taken together, this study serves as a comprehensive characterization of the landscape of metabolic pathway vulnerabilities in cancer cell lines.

scholarly journals Anti-tumor activity of plant extracts against human breast cancer cells are different in monolayer and three-dimensional cell culture screening models: A comparison on 34 extracts

2020 ◽  
Vol 7 (3) ◽  
pp. 3667-3677
Author(s):  
Nhan Lu-Chinh Phan ◽  
Khuong Duy Pham ◽  
Mai Thi-Thanh Nguyen ◽  
Ngoc Kim Phan ◽  
Kiet Dinh Truong ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Plant Extracts ◽  
Breast Cancer Cells ◽  
Three Dimensional ◽  
Culture Model ◽  
Anti Tumor Activity ◽  
Mcf 7

Introduction: The monolayer cell culture model is a popular model for screening anti-tumor activity of plant extracts. However, almost the extracts selected for screening in this model have failed in subsequent animal models. Therefore, there is only about 5 % of candidates from the original thousands of drugs that are screened which ultimately reach clinical trial. This study aimed to compare the differences in anti-tumor activity of 34 plant extracts against breast cancer cells in 2 models of monolayer cell culture (2D) and in three-dimensional (3D) cell culture. Methods: Four breast cancer cell lines (MCF-7, CD44+CD24- MCF-7, VN9, and CD44+CD24- VN9) were used to generate the 2D and 3D models (the 3D model was developed by culturing breast cancer cells in matrigel). The extracts were got from the plant extract library that prepared in the previous study. The anti-tumor activity was evaluated via half inhibitory concentrations( IC50 values). Results: Of the 34 extracts, E12, E7, E5 and E6 of them had an effect on MCF-7, CD44+CD24- MCF-7, VN9 and CD44+CD24- VN9 cells, respectively. The results indicated 10 potentially strong candidates for future drug development targeting hypoxic areas in breast cancer. Conclusion: The 3D culture model exhibited higher resistance to extracts than the 2D culture model. The CD44+CD24- cell population of both VN9 and MCF-7 cell lines showed higher drug resistance than the original cell lines (VN9 and MCF-7).  

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