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Glioblastoma multiforme (GBM) is the most common and lethal type of brain cancer. It is characterized by widespread heterogeneity at the cellular and molecular levels. The detection of this heterogeneity is valuable for accurate diagnosis. Herein, solid-state 20 µm diameter micropore made in thin suspended silicon dioxide membrane is used as cell sensor device. The device relies on a cell's mechano-physical properties as an indicator to differentiate between the subtypes of GBM. A library of GBM cell lines (U251, U87, D54 EGFRviii, and G55) was created by measuring the differences in cell's micropore translocation properties from their distinct electrical profiles. Each GBM subtype has distinct phenotype and this was delineated in their cell translocation behaviors. The library was used to distinguish cells from samples of brain tumor patients. The micropore device accurately profiled GBM patient samples for cell subtypes by comparing data with the GBM library. The micropore approach is simple, can be implemented at low cost and can be used in the clinical setups and operation theaters to detect and identify GBM subtypes from patient samples.
Assuntos
Glioblastoma/patologia , Microtecnologia/instrumentação , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Eletricidade , Humanos , Fenótipo , PorosidadeRESUMO
We present a novel method to trap nanoparticles in double nanohole (DNH) nanoapertures integrated on top of solid-state nanopores (ssNP). The nanoparticles were propelled by an electrophoretic force from the cis towards the trans side of the nanopore but were trapped in the process when they reached the vicinity of the DNH-ssNP interface. The self-induced back action (SIBA) plasmonic force existing between the tips of the DNH opposed the electrophoretic force and enabled simultaneous optical and electrical sensing of a single nanoparticle for seconds. The novel SIBA actuated nanopore electrophoresis (SANE) sensor was fabricated using two-beam GFIS FIB. Firstly, Ne FIB milling was used to create the DNH features and was combined with end pointing to stop milling at the metal-dielectric interface. Subsequently, He FIB was used to drill a 25 nm nanopore through the center of the DNH. Proof of principle experiments to demonstrate the potential utility of the SANE sensor were performed with 20 nm silica and Au nanoparticles. The addition of optical trapping to electrical sensing extended translocation times by four orders of magnitude. The extended electrical measurement times revealed newly observed high frequency charge transients that were attributed to bobbing of the nanoparticle driven by the competing optical and electrical forces. Frequency analysis of this bobbing behavior hinted at the possibility of distinguishing single from multi-particle trapping events. We also discuss how SANE sensor measurement characteristics differ between silica and Au nanoparticles due to differences in their physical properties and how to estimate the charge around a nanoparticle. These measurements show promise for the SANE sensor as an enabling tool for selective detection of biomolecules and quantification of their interactions.
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BACKGROUND AND OBJECTIVE: Detection of metastatic tumor cells is important for early diagnosis and staging of cancer. However, such cells are exceedingly difficult to detect from blood or biopsy samples at the disease onset. It is reported that cancer cells, and especially metastatic tumor cells, show very distinctive morphological behavior compared to their healthy counterparts on aptamer functionalized substrates. The ability to quickly analyze the data and quantify the cell morphology for an instant real-time feedback can certainly contribute to early cancer diagnosis. A supervised machine learning approach is presented for identification and classification of cancer cell gestures for early diagnosis. METHODS: We quantified the morphologically distinct behavior of metastatic cells and their healthy counterparts captured on aptamer-functionalized glass substrates from time-lapse optical micrographs. As a proof of concept, the morphologies of human glioblastoma (hGBM) and astrocyte cells were used. The cells were captured and imaged with an optical microscope. Multiple feature vectors were extracted to quantify and differentiate the complex physical gestures of cancerous and non-cancerous cells. Three different classifier models, Support Vector Machine (SVM), Random Forest Tree (RFT), and Naïve Bayes Classifier (NBC) were trained with the known dataset using machine learning algorithms. The performances of the classifiers were compared for accuracy, precision, and recall measurements using five-fold cross-validation technique. RESULTS: All the classifier models detected the cancer cells with an average accuracy of at least 82%. The NBC performed the best among the three classifiers in terms of Precision (0.91), Recall (0.9), and F1-score (0.89) for the existing dataset. CONCLUSIONS: This paper presents a standalone system built on machine learning techniques for cancer screening based on cell gestures. The system offers rapid, efficient, and novel identification of hGBM brain tumor cells and can be extended to define single cell analysis metrics for many other types of tumor cells.
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Neoplasias Encefálicas/diagnóstico por imagem , Glioblastoma/diagnóstico por imagem , Neoplasias/diagnóstico por imagem , Neoplasias/patologia , Algoritmos , Teorema de Bayes , Neoplasias Encefálicas/diagnóstico , Detecção Precoce de Câncer , Humanos , Processamento de Imagem Assistida por Computador , Modelos Estatísticos , Metástase Neoplásica , Reprodutibilidade dos Testes , Software , Aprendizado de Máquina Supervisionado , Máquina de Vetores de SuporteRESUMO
Metastasis is the major cause of low survival rates among cancer patients. Once cancer cells metastasize, it is extremely difficult to contain the disease. We report on a nanotextured platform for enhanced detection of metastatic cells. We captured metastatic (MDA-MDB-231) and non-metastatic (MCF-7) breast cancer cells on anti-EGFR aptamer modified plane and nanotextured substrates. Metastatic cells were seen to change their morphology at higher rates when captured on nanotextured substrates than on plane substrates. Analysis showed statistically different morphological behaviors of metastatic cells that were very pronounced on the nanotextured substrates. Several distance matrices were calculated to quantify the dissimilarity of cell shape change. Nanotexturing increased the dissimilarity of the metastatic cells and as a result the contrast between metastatic and non-metastatic cells increased. Jaccard distance measurements found that the shape change ratio of the non-metastatic and metastatic cells was enhanced from 1:1.01 to 1:1.81, going from plane to nanotextured substrates. The shape change ratio of the non-metastatic to metastatic cells improved from 1:1.48 to 1:2.19 for the Hausdorff distance and from 1:1.87 to 1:4.69 for the Mahalanobis distance after introducing nanotexture. Distance matrix analysis showed that nanotexture increased the shape change ratios of non-metastatic and metastatic cells. Hence, the detectability of metastatic cells increased. These calculated matrices provided clear and explicit measures to discriminate single cells for their metastatic state on functional nanotextured substrates.
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Aptâmeros de Nucleotídeos/metabolismo , Separação Celular/métodos , Receptores ErbB/metabolismo , Análise de Célula Única/métodos , Alicerces Teciduais , Aptâmeros de Nucleotídeos/síntese química , Linhagem Celular Tumoral , Forma Celular , Feminino , Humanos , Células MCF-7 , Microscopia de Força Atômica , Metástase Neoplásica , Especificidade de Órgãos , Gases em Plasma/química , Propriedades de SuperfícieAssuntos
Calcinose/patologia , Mucosa Gástrica/patologia , Insuficiência Renal Crônica/diagnóstico , Gastropatias/patologia , Neoplasias Gástricas/diagnóstico , Calcinose/diagnóstico , Calcinose/etiologia , Diagnóstico Diferencial , Humanos , Masculino , Diálise Renal , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/terapia , Gastropatias/diagnóstico , Gastropatias/etiologia , Adulto JovemRESUMO
Traumatic injury to the central nervous system is a significant health problem. There is no effective treatment available partly because of the complexity of the system. Implementation of multifunctional micro- and nano-device based combinatorial therapeutics can provide biocompatible and tunable approaches to perform on-demand release of specific drugs. This can help the damaged cells to improve neuronal survival, regeneration of axons, and their reconnection to appropriate targets. Nano-topological features induced rapid cell growth is especially important towards the design of effective platforms to facilitate damaged neural circuit reconstruction. In this study, for the first time, feasibility of neuron-like PC12 cell growth on untreated and easy to prepare nanotextured surfaces has been carried out. The PC12 neuron-like cells were cultured on micro reactive ion etched nanotextured glass coverslips. The effect of nanotextured topology as physical cue for the growth of PC12 cells was observed exclusively, eliminating the possible influence(s) of the enhanced concentration of coated materials on the surface. The cell density was observed to increase by almost 200% on nanotextured coverslips compared to plain coverslips. The morphology study indicated that PC12 cell attachment and growth on the nanotextured substrates did not launch any apoptotic machinery of the cell. Less than 5% cells deformed and depicted condensed nuclei with apoptotic bodies on nanotextured surfaces which is typical for the normal cell handling and culture. Enhanced PC12 cell proliferation by such novel and easy to prepare substrates is not only attractive for neurite outgrowth and guidance, but may be used to increase the affinity of similar cancerous cells (ex: B35 neuroblastoma) and rapid proliferation thereafter-towards the development of combinatorial theranostics to diagnose and treat aggressive cancers like neuroblastoma.
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Proliferação de Células , Animais , Sobrevivência Celular , Neurônios , Células PC12 , RatosRESUMO
Micro- and nanoscale systems have provided means to detect biological targets, such as DNA, proteins, and human cells, at ultrahigh sensitivity. However, these devices suffer from noise in the raw data, which continues to be significant as newer and devices that are more sensitive produce an increasing amount of data that needs to be analyzed. An important dimension that is often discounted in these systems is the ability to quickly process the measured data for an instant feedback. Realizing and developing algorithms for the accurate detection and classification of biological targets in realtime is vital. Toward this end, we describe a supervised machine-learning approach that records single cell events (pulses), computes useful pulse features, and classifies the future patterns into their respective types, such as cancerous/non-cancerous cells based on the training data. The approach detects cells with an accuracy of 70% from the raw data followed by an accurate classification when larger training sets are employed. The parallel implementation of the algorithm on graphics processing unit (GPU) demonstrates a speedup of three to four folds as compared to a serial implementation on an Intel Core i7 processor. This incredibly efficient GPU system is an effort to streamline the analysis of pulse data in an academic setting. This paper presents for the first time ever, a non-commercial technique using a GPU system for realtime analysis, paired with biological cluster targeting analysis.
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Nanoporos , Neoplasias/diagnóstico , Algoritmos , Humanos , Aprendizado de Máquina , Modelos Teóricos , Neoplasias/patologiaRESUMO
Cancer treatment, care, and outcomes are much more effective if started at early stages of the disease. The presence of malignant cancer cells in human samples such as blood or biopsied tissue can be used to reduce overtreatment and underdiagnosis as well as for prognosis monitoring. Reliable quantification of metastatic tumor cells (MTCs) and non-metastatic tumor cells (NMTCs) from human samples can help in cancer staging as well. We report a simple, fast, and reliable approach to identify and quantify metastatic and non-metastatic cancer cells from whole biological samples in a point-of-care manner. The metastatic (MDA MB-231) and non-metastatic (MCF7) breast cancer cells were pushed through a solid-state micropore made in a 200 nm thin SiO2 membrane while measuring current across the micropore. The cells generated very distinctive translocation profiles. The translocation differences stemmed from their peculiar mechanophysical properties. The detection efficiency of the device for each type of tumor cells was â¼75%. MTCs showed faster translocation (36%) and 34% less pore blockage than NMTCs. The micropore approach is simple, exact, and quantitative for metastatic cell detection in a lab-on-a chip setting, without the need for any preprocessing of the sample.
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Separação Celular/instrumentação , Análise Serial de Tecidos/instrumentação , Neoplasias da Mama/patologia , Movimento Celular , Humanos , Células MCF-7 , Metástase Neoplásica , PorosidadeRESUMO
Tumor cells are malignant derivatives of normal cells. There are characteristic differences in the mechanophysical properties of normal and tumor cells, and these differences stem from the changes that occur in the cell cytoskeleton during cancer progression. There is a need for viable whole blood processing techniques for rapid and reliable tumor cell detection that do not require tagging. Micropore biosensors have previously been used to differentiate tumor cells from normal cells and we have used a micropore-based electromechanical transducer to differentiate one type of tumor cells from the other types. This device generated electrical signals that were characteristic of the cell properties. Three non-small cell lung cancer (NSCLC) cell lines, NCl-H1155, A549 and NCI-H460, were successfully differentiated. NCI-H1155, due to their comparatively smaller size, were found to be the quickest in translocating through the micropore. Their translocation through a 15 µm micropore caused electrical pulses with an average translocation time of 101 ± 9.4 µs and an average peak amplitude of 3.71 ± 0.42 µA, whereas translocation of A549 and NCI-H460 caused pulses with average translocation times of 126 ± 17.9 µs and 148 ± 13.7 µs and average peak amplitudes of 4.58 ± 0.61 µA and 5.27 ± 0.66 µA, respectively. This transformation of the differences in cell properties into differences in the electrical profiles (i.e. the differences in peak amplitudes and translocation times) with this electromechanical transducer is a quantitative way to differentiate these lung cancer cells. The solid-state micropore device processed whole biological samples without any pre-processing requirements and is thus ideal for point-of-care applications.
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Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Detecção Precoce de Câncer/instrumentação , Neoplasias Pulmonares/diagnóstico , Linhagem Celular Tumoral , Tamanho Celular , Detecção Precoce de Câncer/métodos , Humanos , Dispositivos Lab-On-A-Chip , Fenômenos MecânicosRESUMO
Microfluidic channels have been implemented to detect cancer cells from blood using electrical measurement of each single cell from the sample. Every cell provided characteristic current profile based on its mechano-physical properties. Cancer cells not only showed higher translocation time and peak amplitude compared to blood cells, their pulse shape was also distinctively different. Prevalent microfluidic channels are plain but we created nanotexture on the channel walls using micro reactive ion etching (micro-RIE). The translocation behaviors of the metastatic renal cancer cells through plain and nanotextured PDMS microchannels showed clear differences. Nanotexture enhanced the cell-surface interactions and more than 50% tumor cells exhibited slower translocation through nanotextured channels compared to plain devices. On the other hand, most of the blood cells had very similar characteristics in both channels. Only 7.63% blood cells had slower translocation in nanotextured microchannels. The tumor cell detection efficiency from whole blood increased by 14% in nanotextured microchannels compared to plain channels. This interesting effect of nanotexture on translocation behavior of tumor cells is important for the early detection of cancer.
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Técnicas Analíticas Microfluídicas , Microfluídica/métodos , Células Neoplásicas Circulantes , Análise de Célula Única/métodos , Animais , Células Sanguíneas , Neoplasias Encefálicas/secundário , Humanos , Neoplasias Renais/patologia , Técnicas Analíticas Microfluídicas/instrumentação , Microfluídica/instrumentação , Neoplasias/diagnóstico , RatosRESUMO
Detection of circulating tumor cells (CTCs) in the early stages of cancer is a great challenge because of their exceedingly small concentration. There are only a few approaches sensitive enough to differentiate tumor cells from the plethora of other cells in a sample like blood. In order to detect CTCs, several antibodies and aptamers have already shown high affinity. Nanotexture can be used to mimic basement membrane to further enhance this affinity. This article reports an approach to fabricate nanotextured polydimethylsiloxane (PDMS) substrates using micro reactive ion etching (micro-RIE). Three recipes were used to prepare nanotextured PDMS using oxygen and carbon tetrafluoride. Micro-RIE provided better control on surface properties. Nanotexturing improved the affinity of PDMS surfaces to capture cancer cells using surface immobilized aptamers against cell membrane overexpressed with epidermal growth factor receptors. In all cases, nanotexture of PDMS increased the effective surface area by creating nanoscale roughness on the surface. Nanotexture also enhanced the growth rate of cultured cells compared to plain surfaces. A comparison among the three nanotextured surfaces demonstrated an almost linear relationship between the surface roughness and density of captured tumor cells. The nanotextured PDMS mimicked biophysical environments for cells to grow faster. This can have many implications in microfluidic platforms used for cell handling.
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Neoplasias Encefálicas/diagnóstico , Separação Celular/métodos , Glioblastoma/diagnóstico , Nanotecnologia/métodos , Aptâmeros de Nucleotídeos/metabolismo , Astrócitos/citologia , Linhagem Celular Tumoral , Separação Celular/instrumentação , Dimetilpolisiloxanos/química , Humanos , Nanoestruturas/química , Nanotecnologia/instrumentação , Células Neoplásicas Circulantes/metabolismoRESUMO
Aptamers are single-stranded DNA or RNA oligomers, identified from a random sequence pool, with the ability to form unique and versatile tertiary structures that bind to cognate molecules with superior specificity. Their small size, excellent chemical stability and low immunogenicity enable them to rival antibodies in cancer imaging and therapy applications. Their facile chemical synthesis, versatility in structural design and engineering, and the ability for site-specific modifications with functional moieties make aptamers excellent recognition motifs for cancer biomarker discovery and detection. Moreover, aptamers can be selected or engineered to regulate cancer protein functions, as well as to guide anti-cancer drug design or screening. This review summarizes their applications in cancer, including cancer biomarker discovery and detection, cancer imaging, cancer therapy, and anti-cancer drug discovery. Although relevant applications are relatively new, the significant progress achieved has demonstrated that aptamers can be promising players in cancer research.
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Aptâmeros de Nucleotídeos/química , Neoplasias/diagnóstico , Neoplasias/terapia , Antineoplásicos/uso terapêutico , Biomarcadores , Desenho de Fármacos , HumanosRESUMO
In this paper, we report a one-step tumor cell detection approach based on the dynamic morphological behavior tracking of cancer cells on a ligand modified surface. Every cell on the surface was tracked in real time for several minutes immediately after seeding until these were finally attached. Cancer cells were found to be very active in the aptamer microenvironment, changing their shapes rapidly from spherical to semi-elliptical, with much flatter spread and extending pseudopods at regular intervals. When incubated on a functionalized surface, the balancing forces between cell surface molecules and the surface-bound aptamers, together with the flexibility of the membranes, caused cells to show these distinct dynamic activities and variations in their morphologies. On the other hand, healthy cells remained distinguishingly inactive on the surface over the same period. The quantitative image analysis of cell morphologies provided feature vectors that were statistically distinct between normal and cancer cells.
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This paper presents a simple approach to create a two-tiered surface for superior cancer cell isolation. The idea is inspired by the interactions of cells with a nanotextured basement membrane. The texture mimicked the extracellular matrix and basement membrane for superior target cell adhesion. Prepared micro+nanotextured surfaces showed enhanced cell capture. Preparation of the two-tiered surface was done using micro- and nanotexturing and was easily reproducible. It has been shown before that the larger surface area of a nanotextured surface assists the cell's attachment through surface-anchored ligands. Taking it a step further, ligand functionalized two-level micro+nanotextured surfaces improved the sensitivity of the cancer cell isolation over simple flat nanotexturing. The isolation efficiency increased by 208% compared to the surface with a single-level nanotexture. The two-tiered surface was compatible with previously reported nanotextured devices used for cancer cell isolation. Micro-texture on the glass surface was created using simple sand gritting, followed by reactive ion etching (RIE) of the entire surface. The approach could create large surface areas within a short time while maintaining superior cell isolation efficiency.
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Separação Celular , Glioblastoma/diagnóstico , Nanoestruturas/química , Membrana Basal , Adesão Celular , Matriz Extracelular , Humanos , LigantesRESUMO
Early stage detection and precise quantification of circulating tumor cells (CTCs) in the peripheral blood of cancer patients are important for early diagnosis. Early diagnosis improves the effectiveness of the therapy and results in better prognosis. Several techniques have been used for CTC detection but are limited by their need for dye tagging, low throughput and lack of statistical reliability at single cell level. Solid-state micropores can characterize each cell in a sample providing interesting information about cellular populations. We report a multi-channel device which utilized solid-state micropores array assembly for simultaneous measurement of cell translocation. This increased the throughput of measurement and as the cells passed the micropores, tumor cells showed distinctive current blockade pulses, when compared to leukocytes. The ionic current across each micropore channel was continuously monitored and recorded. The measurement system not only increased throughput but also provided on-chip cross-relation. The whole blood was lysed to get rid of red blood cells, so the blood dilution was not needed. The approach facilitated faster processing of blood samples with tumor cell detection efficiency of about 70%. The design provided a simple and inexpensive method for rapid and reliable detection of tumor cells without any cell staining or surface functionalization. The device can also be used for high throughput electrophysiological analysis of other cell types.
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Técnicas Biossensoriais/instrumentação , Filtros Microporos , Neoplasias/sangue , Neoplasias/diagnóstico , Células Neoplásicas Circulantes/patologia , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/secundário , Detecção Precoce de Câncer , Desenho de Equipamento , Hemólise , Humanos , Neoplasias Renais/patologiaRESUMO
Biomarker-binding nucleotide sequences, like aptamers, have gained recent attention in cancer cell isolation and detection works. Self-assembly and 3D conformation of aptamers enable them to selectively capture and bind diseased cells and related biomarkers. One mode of utilizing such an extraordinary selective property of the aptamers is by grafting these in nanopores. Coating the inside walls of the nanopore with biomarker specific ligands, like DNA, changes the statistics of the dynamic translocation events. When the target protein passes through the nanopore, it interacts with ligand coated inside the nanopore, and the process alters the overall potential energy profile which is essentially specific to the protein detected. The fundamental goal in this process is to ensure that these detection motifs hold their structure and functionality under applied electric field and experimental conditions. We report here all-atom molecular dynamics simulations of the effects of external electric field on the 3D conformation of such DNA structures. The simulations demonstrate how the grafted moieties affect the translocation time, velocity, and detection frequency of the target molecule. We also investigated a novel case of protein translocation, where DNA is prebound to the protein. As model, a thrombin-specific G-quartet and thrombin pair was used for this study.
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Aptâmeros de Nucleotídeos/metabolismo , DNA de Cadeia Simples/química , Nanoporos/ultraestrutura , Trombina/metabolismo , Aptâmeros de Nucleotídeos/química , Sequência de Bases , Técnicas Eletroquímicas/instrumentação , Desenho de Equipamento , Humanos , Simulação de Dinâmica Molecular , Conformação de Ácido Nucleico , Conformação Proteica , Transporte Proteico , Trombina/análiseRESUMO
Proteomics information of cancer has shown that abnormalities at the levels of growth factors, receptors, intracellular mediators and transcription factors play major role in the disease progression. We report a directly quantifiable approach to measure tumor cell behavior on functionalized chips. The chips were functionalized with aptamer molecules that were selective against epidermal growth factor receptor (EGFR), a commonly overexpressed cancer biomarker. The chip-bound aptamer selectively isolated tumor cells from cell mixture samples. The isolated cells were thus bound to the chip surface. However, some normal cells also got captured on the surface. The selectivity and sensitivity of tumor isolation changed when the surface of the chip was chemically treated to create nanoscale texture. The captured cancer cells showed distinctly different behavior on the surface of the chip than that for the normal cells. The behavior quantification can serve as a novel modality to detect cancer cells from simple samples like blood, saliva or urine.
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Biomarcadores Tumorais/análise , Separação Celular/instrumentação , Análise em Microsséries/instrumentação , Neoplasias/química , Células Neoplásicas Circulantes/química , Biomarcadores Tumorais/química , Humanos , Nanotecnologia/instrumentação , Neoplasias/metabolismo , Células Neoplásicas Circulantes/metabolismoRESUMO
Rare cells already have become established indicators for disease diagnosis, to help track prognosis, and in developing personalized therapy. Numerous techniques have been developed to effectively and specifically detect and sort rare cells and cell isolation techniques have gained much attention among researchers in the last few decades. Recent developments in nanotechnologies and microfluidics have been used with great promise towards these goals. The research emphasis has also shifted from simple detection with microfluidic devices to comprehensive isolation, collection and subsequent analysis with integrated and automated systems. The first challenge in post-isolation analysis is cell detachment from substrates, while keeping cells viable and unperturbed. In this review, various methods used for cell detachments are discussed. For effective cell sorting, the detachment is identified as critical criteria for selecting substrates and methods.
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Fenômenos Fisiológicos Celulares , Separação Celular , Sobrevivência Celular , Técnicas Analíticas Microfluídicas , Animais , Células COS , Adesão Celular , Células Cultivadas , Chlorocebus aethiops , Humanos , Camundongos , PolímerosRESUMO
Tumor cells depict two deviant tendencies; over-proliferation and vigorous migration. A tapered channel device is designed and fabricated for in vitro studies. We report inhibited proliferation and migration of human glioblastoma (hGBM) cells when exposed to an aptamer that is known to bind epidermal growth factor receptors (EGFR). The device is integrated with controlled ambient and microscope for providing real-time and quantitative characterization of the tumor cell behavior. The results show that hGBM cells loose proliferation and motility when exposed to the anti-EGFR aptamer. The aptamer directly inhibits and blocks EGF-induced EGFR phosphorylation. This also reduces the ability of cells to remodel their internal structure for invasion through narrow constrictions. This provides a framework for possible studies on efficacy of other inhibiting molecules.
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Biomimética/métodos , Movimento Celular , Glioblastoma/patologia , Técnicas Analíticas Microfluídicas/métodos , Aptâmeros de Nucleotídeos/metabolismo , Divisão Celular , Proliferação de Células , Receptores ErbB/metabolismo , Humanos , Fosforilação , Transdução de SinaisRESUMO
Cell adhesion, morphology and growth are influenced by surface topography at nano and micrometer scales. Nano-textured surfaces are prepared using photolithography, plasma etching and long polymer chemical etching which are cost prohibitive and require specialized equipment. This article demonstrates a simple approach to synthesize nano-textured scaffolds from chicken eggshells. Varieties of pattern are made on the eggshells like micro-needle forests and nanopores, giving very uniform nano-textures to the surfaces. The surfaces are characterized for chemical composition and crystal phase. The novel patterns are transferred to PDMS surfaces and the nano-textured PDMS surfaces are used to study the effect of texturing on human fibroblast cell growth and attachment. The effects of surface topographies, along with laminin coating on cell cultures, are also studied. We find an exciting phenomenon that the initial seeding density of the fibroblast cells affects the influence of the nano-texturing on cell growth. These nano-textured surfaces give 16 times more fibroblast growth when compared to flat PDMS surfaces. The novel nano-textured patterns also double the laminin adsorption on PDMS.