An Electrical Impedance-Based Assay to Examine Functions of Various Placental Cell Types In Vitro

Chollangi, Tejasvy; Clabault, Hélène; Thibeault, Andrée Anne Hudon; Yong, Hannah Ee Juen; Narula, Shagun; Menkhorst, Ellen; Sanderson, J. Thomas; Vaillancourt, Cathy; Murthi, Padma

doi:10.1007/978-1-4939-7498-6_20

Cited by 3 publications

(2 citation statements)

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“…The xCELLigence RTCA eSight machine (Agilent) was used to obtain real-time impedance-based measurements of cell adhesion and proliferation (Chollangi et al, 2018). The user can set a temporal resolution (how frequent the impedance value is measured) in which an adimensional value called "cell index" is measured.…”

Section: Esightmentioning

confidence: 99%

Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer

Pisano

Pierini

et al. 2020

Front. Cell Dev. Biol.

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Exosomes are physiologically secreted nanoparticles recently established as natural delivery systems involved in cell-to-cell communication and content exchange. Due to their inherent targeting potential, exosomes are currently being harnessed for the development of anti-cancer therapeutics. Clinical trials evaluating their effectiveness are demonstrating safety and promising outcomes. However, challenging largescale production, isolation, modification and purification of exosomes are current limitations for the use of naturally occurring exosomes in the clinic. Exosome mimetics hold the promise to improve the delivery of bioactive molecules with therapeutic efficacy, while achieving scalability and increasing bioavailability. In this study, we propose the development of Immune Derived Exosome Mimetics (IDEM) as a scalable approach to target and defeat ovarian cancer cells. IDEM were fabricated from monocytic cells by combining sequential filtration steps through filter membranes with different porosity and size exclusion chromatography columns. The physiochemical and molecular characteristics of IDEM were compared to those of natural exosomes (EXO). Nanoparticle Tracking Analysis confirmed a 2.48-fold increase in the IDEM production yields compared to EXO, with similar exosomal markers profiles (CD81, CD63) as demonstrated by flow cytometry and ELISA. To exploit the prospective of IDEM to deliver chemotherapeutics, doxorubicin (DOXO) was used as a model drug. IDEM showed higher encapsulation efficiency and drug release over time compared to EXO. The uptake of both formulations by SKOV-3 ovarian cancer cells was assessed by confocal microscopy and flow cytometry, showing an incremental drug uptake over time. The analysis of the cytotoxic and apoptotic effect of DOXOloaded nanoparticles both in 2D and 3D culture systems proved IDEM as a more efficient system as compared to free DOXO, unraveling the advantage of IDEM in reducing side-effects while increasing cytotoxicity of targeted cells, by delivering smaller amount of the chemotherapeutic agent. The high yields of IDEM obtained compared to natural exosomes together with the time-effectiveness and reproducibility of their production method make this approach potentially exploitable for clinical applications.

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Section: Esightmentioning

confidence: 99%

Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer

Pisano

Pierini

et al. 2020

Front. Cell Dev. Biol.

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“…This principle has been used to develop different real-time migration and invasion assay devices. For example, the commercial xCELLigence system was used for migration and invasion study of various cell types including placental and endothelial cells (21,22). EIS was used in a simple invasion assay by integrating the electrodes in a transwell-inspired device (23).…”

Section: Background and Rationalementioning

confidence: 99%

Development of paper-based 3D cell culture devices and real-time biosensing applications

Pupinyo

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Due to its micro fibrous pores, paper has gained traction in the use as a scaffold for 3D cell culture.�To study melanoma mechanism, screening the amount of melanin production and invasion study are major keys for drug development. In this work, we developed paper-based devices for culturing melanoma cells. Through the development process, we obtained the optimal parameters, which were applied for 3 different projects; 1) Anti-melanogenic effect screening, 2) Impedance-based E-screen assay, and 3) A membrane insert for real-time invasion assay. As a result of melanin production from melanocytes, the cells can be easily observed after a few days of culturing due to their black color. Fortunately, this color change can easily be visualized when melanocyte is cultured in the paper due to the white color of the paper. The paper was then applied for anti-melanogenic screening of the natural compounds with short analysis time and easy steps. In order to develop the real-time invasion assay, we primarily study the impedance characteristics of cells. The obtained impedance parameters were applied for real-time estrogen screening as a parallel project. Based on the estrogen receptor on breast cancer cells, the binding of estrogen can increase cell proliferation, resulting in the increased impedance magnitude. The estrogenic effect of the test compounds, Bisphenol-A and Irgarol 1051, were�then monitored in real time. After that, we applied the obtained impedance parameters for the development of real-time invasion assay device using paper as a membrane insert. The concept was based on the blockage of invasive cells within paper pores, resulting in the increased impedance magnitude. The device was applied for real-time invasion study of melanoma cells under the treatment of IGF-1.��Melanoma invasion could be tracked within 7 h after IGF-1 treatment.�Thus, the invasion of melanoma cells was less likely affected by cell proliferation.�

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