Single-cell assays

Ryan, Declan; Ren, Kangning; Wu, Hongkai

doi:10.1063/1.3574448

Cited by 30 publications

(21 citation statements)

References 40 publications

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“…The analysis of biochemical contents, such as protein,164 DNA and RNA,165 in single cells, is critical for the understanding of cellular processes 166. Unlike conventional analysis for a batch of cells, single‐cell analysis could avoid the loss of information associated with ensemble averaging, and thus reflect important variations in a cellular population.…”

Section: Single Cell Analysismentioning

confidence: 99%

Microfluidics for Manipulating Cells

Zheng

Sun

et al. 2012

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Microfluidics, a toolbox comprising methods for precise manipulation of fluids at small length scales (micrometers to millimeters), has become useful for manipulating cells. Its uses range from dynamic management of cellular interactions to high-throughput screening of cells, and to precise analysis of chemical contents in single cells. Microfluidics demonstrates a completely new perspective and an excellent practical way to manipulate cells for solving various needs in biology and medicine. This review introduces and comments on recent achievements and challenges of using microfluidics to manipulate and analyze cells. It is believed that microfluidics will assume an even greater role in the mechanistic understanding of cell biology and, eventually, in clinical applications.

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Section: Single Cell Analysismentioning

confidence: 99%

Microfluidics for Manipulating Cells

Zheng

Sun

et al. 2012

Small

180

130

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“…Moreover, profiling does not provide information on protein-protein interactions (Hunter, 2000 ; Celis and Gromov, 2003 ), and most critically, nor does it fully account for the extreme clonal heterogeneity recently revealed (Barbieri et al, 2013 ). In this regard, the possibility to functionally test treatment protocols directly on patient tumor cells, before therapy implementation, in an “ in vivo mimicking” environment, and preferably by utilizing single cell assays (Ryan et al, 2011 ), or at a minimum by performing studies at the clonal level (Sabaawy, 2014 ), would definitely address these limitations. Here, we present the key strengths, weaknesses, and possible applications of the 3D organoid models into the biomedical field, with a particular emphasis on the prostate cancer (PCa) therapeutic arena.…”

mentioning

confidence: 99%

Personalized Medicine Approaches in Prostate Cancer Employing Patient Derived 3D Organoids and Humanized Mice

Bartucci

Ferrari

Kim

et al. 2016

Front. Cell Dev. Biol.

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Prostate cancer (PCa) is the most common malignancy and the second most common cause of cancer death in Western men. Despite its prevalence, PCa has proven very difficult to propagate in vitro. PCa represents a complex organ-like multicellular structure maintained by the dynamic interaction of tumoral cells with parenchymal stroma, endothelial and immune cells, and components of the extracellular matrix (ECM). The lack of PCa models that recapitulate this intricate system has hampered progress toward understanding disease progression and lackluster therapeutic responses. Tissue slices, monolayer cultures and genetically engineered mouse models (GEMM) fail to mimic the complexities of the PCa microenvironment or reproduce the diverse mechanisms of therapy resistance. Moreover, patient derived xenografts (PDXs) are expensive, time consuming, difficult to establish for prostate cancer, lack immune cell-tumor regulation, and often tumors undergo selective engraftments. Here, we describe an interdisciplinary approach using primary PCa and tumor initiating cells (TICs), three-dimensional (3D) tissue engineering, genetic and morphometric profiling, and humanized mice to generate patient-derived organoids for examining personalized therapeutic responses in vitro and in mice co-engrafted with a human immune system (HIS), employing adaptive T-cell- and chimeric antigen receptor- (CAR) immunotherapy. The development of patient specific therapies targeting the vulnerabilities of cancer, when combined with antiproliferative and immunotherapy approaches could help to achieve the full transformative power of cancer precision medicine.

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“…Studies done on large heterogeneous cell samples yield an average view which can obscure detection and examination of important minority cell types (Holmes and Al-Rubeai 1999; Love et al 2006; Jin et al 2009; Gong et al 2010; Nikkah et al 2011; Ryan et al 2011). In developing assays for single cell screening using micro-well arrays it is necessary to identify and fully characterize the factors that determine cell seeding efficiency and distribution statistics.…”

Section: Discussionmentioning

confidence: 99%

Characterization of cell seeding and specific capture of B cells in microbubble well arrays

Jones

Kobie

DeLouise

2013

Biomed Microdevices

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Development of micro-well array systems for use in high-throughput screening of rare cells requires a detailed understanding of the factors that impact the specific capture of cells in wells and the distribution statistics of the number of cells deposited into wells. In this study we investigate the development of microbubble (MB) well array technology for sorting antigen-specific B-cells. Using Poisson statistics we delineate the important role that the fractional area of MB well opening and the cell seeding density have on determining cell seeding distribution in wells. The unique architecture of the MB well hinders captured cells from escaping the well and provides a unique microenvironmental niche that enables media changes as needed for extended cell culture. Using cell lines and primary B and T cells isolated from human peripheral blood we demonstrate the use of affinity capture agents coated in the MB wells to enrich for the selective capture of B cells. Important differences were noted in the efficacy of bovine serum albumin to block the nonspecific adsorption of primary cells relative to cell lines as well as the efficacy of the capture coatings using mixed primary B and T cells samples. These results emphasize the importance of using primary cells in technology development and suggest the need to utilize B cell capture agents that are insensitive to cell activation.

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Single-cell assays

Cited by 30 publications

References 40 publications

Microfluidics for Manipulating Cells

Microfluidics for Manipulating Cells

Personalized Medicine Approaches in Prostate Cancer Employing Patient Derived 3D Organoids and Humanized Mice

Characterization of cell seeding and specific capture of B cells in microbubble well arrays

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