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      Microfabricated Electrochemical Cell-Based Biosensors for Analysis of Living Cells In Vitro

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          Abstract

          Cellular biochemical parameters can be used to reveal the physiological and functional information of various cells. Due to demonstrated high accuracy and non-invasiveness, electrochemical detection methods have been used for cell-based investigation. When combined with improved biosensor design and advanced measurement systems, the on-line biochemical analysis of living cells in vitro has been applied for biological mechanism study, drug screening and even environmental monitoring. In recent decades, new types of miniaturized electrochemical biosensor are emerging with the development of microfabrication technology. This review aims to give an overview of the microfabricated electrochemical cell-based biosensors, such as microelectrode arrays (MEA), the electric cell-substrate impedance sensing (ECIS) technique, and the light addressable potentiometric sensor (LAPS). The details in their working principles, measurement systems, and applications in cell monitoring are covered. Driven by the need for high throughput and multi-parameter detection proposed by biomedicine, the development trends of electrochemical cell-based biosensors are also introduced, including newly developed integrated biosensors, and the application of nanotechnology and microfluidic technology.

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          Oxidized redox state of glutathione in the endoplasmic reticulum.

          C. Hwang, A Sinskey, H Lodish (1992)
          The redox state of the endoplasmic reticulum (ER) was measured with the peptide N-Acetyl-Asn-Tyr-Thr-Cys-NH2. The peptide diffused across cellular membranes; some became glycosylated and thus trapped within the secretory pathway, and its cysteine residue underwent reversible thiol-disulfide exchanges with the surrounding redox buffer. Glycosylated peptides from cells were disulfide-linked to glutathione, indicating that glutathione is the major redox buffer in the secretory pathway. The redox state of the secretory pathway was more oxidative than that of the cytosol; the ratio of reduced glutathione to the disulfide form (GSH/GSSG) within the secretory pathway ranged from 1:1 to 3:1, whereas the overall cellular GSH/GSSG ratio ranged from 30:1 to 100:1. Cytosolic glutathione was also transported into the lumen of microsomes in a cell-free system. Although how the ER maintains an oxidative environment is not known, these results suggest that the demonstrated preferential transport of GSSG compared to GSH into the ER lumen may contribute to this redox compartmentation.
          Article 0 comments Cited 288 times – based on 0 reviews     Review now
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            Label-free immunodetection with CMOS-compatible semiconducting nanowires.

            Eric Stern, James F Klemic, David Routenberg … (2007)
            Semiconducting nanowires have the potential to function as highly sensitive and selective sensors for the label-free detection of low concentrations of pathogenic microorganisms. Successful solution-phase nanowire sensing has been demonstrated for ions, small molecules, proteins, DNA and viruses; however, 'bottom-up' nanowires (or similarly configured carbon nanotubes) used for these demonstrations require hybrid fabrication schemes, which result in severe integration issues that have hindered widespread application. Alternative 'top-down' fabrication methods of nanowire-like devices produce disappointing performance because of process-induced material and device degradation. Here we report an approach that uses complementary metal oxide semiconductor (CMOS) field effect transistor compatible technology and hence demonstrate the specific label-free detection of below 100 femtomolar concentrations of antibodies as well as real-time monitoring of the cellular immune response. This approach eliminates the need for hybrid methods and enables system-scale integration of these sensors with signal processing and information systems. Additionally, the ability to monitor antibody binding and sense the cellular immune response in real time with readily available technology should facilitate widespread diagnostic applications.
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              Concomitant activation of the JAK/STAT, PI3K/AKT, and ERK signaling is involved in leptin-mediated promotion of invasion and migration of hepatocellular carcinoma cells.

              Dipali Sharma, Didier Merlin, Kirti N Saxena … (2007)
              Various epidemiologic studies have shown that obesity is associated with hepatocellular carcinoma. Leptin, the key player in the regulation of energy balance and body weight control, also acts as a growth factor on certain organs in both normal and disease states. It is plausible that leptin acts to promote hepatocellular carcinogenesis directly affecting malignant properties of liver cancer cells. However, a direct role for leptin in hepatocellular carcinoma has not been shown. In this study, we analyzed the role of leptin and the mechanism(s) underlying its action in hepatocellular carcinoma cells, which express both short and long isoforms of leptin receptors. Treatment with leptin resulted in increased proliferation of both HepG2 and Huh7 cells and involves activation of signal transducers and activators of transcription 3 (STAT3), AKT, and extracellular signal-regulated kinase (ERK) signaling pathways. Leptin-induced phosphorylation of ERK and AKT was dependent on Janus-activated kinase (JAK)/STAT activation. Intriguingly, we also found that leptin potently induces invasion of hepatocellular carcinoma cells in Matrigel invasion and electric cell-substrate impedance-sensing assays. Leptin-stimulated invasion was effectively blocked by pharmacologic inhibitors of JAK/STAT and, to a lesser extent, by ERK and phosphatidylinositol 3-kinase (PI3K) inhibition. Importantly, leptin also induced the migration of both HepG2 and Huh7 cells on fibronectin matrix. Inhibition of JAK/STAT, ERK, and PI3K activation using pharmacologic inhibitors effectively blocked leptin-induced migration of HepG2 and Huh7 cells. Taken together, these data indicate that leptin promotes hepatocellular carcinoma growth, invasiveness, and migration and implicate the JAK/STAT pathway as a critical mediator of leptin action. Our findings have potential clinical implications for hepatocellular carcinoma progression in obese patients.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Biosensors (Basel)
                Journal ID (iso-abbrev): Biosensors (Basel)
                Journal ID (publisher-id): biosensors
                Title: Biosensors
                Publisher: MDPI
                ISSN (Electronic): 2079-6374
                Publication date (Electronic): 25 April 2012
                Publication date Collection: June 2012
                Volume: 2
                Issue: 2
                Pages: 127-170
                Affiliations
                Biosensor National Special Lab, Key Lab for Biomedical Engineering of Ministry of Education, Department of Biomedical Engineering, Zheda Road No. 38, Zhejiang University, Hangzhou 310027, China; E-Mail: wangjun-47@ 123456163.com (J.W.); yebear@ 123456zju.edu.cn (C.W.); zjubmehuning@ 123456gmail.com (N.H.); zhoujiecomeon@ 123456163.com (J.Z.); dlp032506@ 123456hotmail.com (L.D.)
                Author notes
                [* ]Author to whom correspondence should be addressed; E-Mail: cnpwang@ 123456zju.edu.cn ; Tel./Fax: +86-571-8795-2832.
                Article
                Publisher ID: biosensors-02-00127
                DOI: 10.3390/bios2020127
                PMC ID: 4263572
                SO-VID: 74b9973b-61ab-4343-bfb5-01fea865cd75
                Copyright © © 2012 by the authors; licensee MDPI, Basel, Switzerland.
                License:

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

                History
                Date received : 02 March 2012
                Date revision received : 02 April 2012
                Date accepted : 19 April 2012
                Categories
                Subject: Review

                Keywords: electrochemical biosensor,cell-based biosensor,microfabrication,in vitro monitoring
                Data availability:
                Keywords: electrochemical biosensor, cell-based biosensor, microfabrication, in vitro monitoring

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