scholarly journals Labeling of multiple cell lines using a new iron oxide agent for cell tracking by MRI

2011 ◽  
Vol 6 (6) ◽  
pp. 514-522 ◽  
Author(s):  
C. McFadden ◽  
C. L. Mallett ◽  
P. J. Foster
Keyword(s):  
Iron Oxide ◽  
Cell Lines ◽  
Cell Tracking ◽  
Multiple Cell

scholarly journals Combinatorial CRISPR screen identifies fitness effects of gene paralogues

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Nicola A. Thompson ◽  
Marco Ranzani ◽  
Louise van der Weyden ◽  
Vivek Iyer ◽  
Victoria Offord ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Cell Lines ◽  
Unknown Function ◽  
Single Copy ◽  
Genetic Redundancy ◽  
Synthetic Lethal ◽  
Fitness Effects ◽  
Synthetic Lethal Interactions ◽  
Crispr Screen ◽  
Multiple Cell

AbstractGenetic redundancy has evolved as a way for human cells to survive the loss of genes that are single copy and essential in other organisms, but also allows tumours to survive despite having highly rearranged genomes. In this study we CRISPR screen 1191 gene pairs, including paralogues and known and predicted synthetic lethal interactions to identify 105 gene combinations whose co-disruption results in a loss of cellular fitness. 27 pairs influence fitness across multiple cell lines including the paralogues FAM50A/FAM50B, two genes of unknown function. Silencing of FAM50B occurs across a range of tumour types and in this context disruption of FAM50A reduces cellular fitness whilst promoting micronucleus formation and extensive perturbation of transcriptional programmes. Our studies reveal the fitness effects of FAM50A/FAM50B in cancer cells.

scholarly journals 2173 RNA-nanoparticles to enhance and track dendritic cell migration

2018 ◽  
Vol 2 (S1) ◽  
pp. 26-26
Author(s):  
Adam J. Grippin ◽  
Elias J. Sayour ◽  
Brandon Wummer ◽  
Adam Monsalve ◽  
Tyler Wildes ◽  
...  
Keyword(s):  
Iron Oxide ◽  
Lymph Nodes ◽  
Cell Tracking ◽  
Ex Vivo ◽  
Imaging Modality ◽  
Cationic Lipids ◽  
Widespread Application ◽  
Vaccine Site ◽  
Stimulatory Capacity

OBJECTIVES/SPECIFIC AIMS: Despite aggressive chemotherapy, surgical resection, and radiation therapy, glioblastoma remains almost universally fatal. In a pilot, randomized, and blinded clinical trial, we recently demonstrated that administration of RNA-loaded DC vaccines was associated with significantly improved progression-free and overall survival in patients with glioblastoma (Mitchell et al., Nature, 2015). Furthermore, clinical outcomes correlated with DC migration to vaccine-site draining lymph nodes measured by Indium-111 labeling of RNA-loaded DCs and SPECT/CT imaging. Although these studies demonstrated that tracking DC migration may be an important clinical biomarker for response to DC vaccination, the complexity and regulatory requirements associated with nuclear labelling to track DC migration limits widespread application of this technique. We have therefore developed RNA-loaded magnetic nanoparticles (RNA-NPs) to enhance DC migration to LNs and track that migration with a widely available imaging modality (i.e., MRI). METHODS/STUDY POPULATION: Cationic liposomes were loaded with iron oxide nanoparticles with or without cholesterol. The resulting nanoparticles were complexed with RNA and used to transfect DCs ex vivo. RNA-NP-loaded DsRed+ DCs were then injected intradermally into mice and tracked noninvasively with T2-weighted 11T MRI before excision and quantification with flow cytometry. RESULTS/ANTICIPATED RESULTS: In vitro experiments demonstrate that iron oxide loading does not reduce RNA-NP-mediated transfection of DCs. Additionally, replacement of cationic lipids with cholesterol increased RNA-NP transfection of the DC2.4 cell line and enhanced the T cell stimulatory capacity of treated bone marrow-derived dendritic cells (BMDCs). Compared to electroporation, RNA-NPs enhanced DC migration to lymph nodes and reduced T2 MRI intensity in DC-bearing lymph nodes. DISCUSSION/SIGNIFICANCE OF IMPACT: This data suggests that iron oxide-loaded RNA-NPs enable noninvasive cell tracking with MRI and enhance DC migration to lymph nodes. We have further shown that inclusion of cholesterol in RNA-NPs augments the stimulatory capacity of transfected DCs. Future work will consider effects of RNA-NPs on antitumor immune responses and the utility of MRI-detected DC migration as a biomarker of vaccine efficacy.

Efficient and Rapid Labeling of Transplanted Cell Populations with Superparamagnetic Iron Oxide Nanoparticles Using Cell Surface Chemical Biotinylation for in Vivo Monitoring by MRI

2010 ◽  
Vol 19 (4) ◽  
pp. 419-429 ◽  
Author(s):  
Po-Wah So ◽  
Tammy Kalber ◽  
David Hunt ◽  
Michael Farquharson ◽  
Alia Al-Ebraheem ◽  
...  
Keyword(s):  
Iron Oxide ◽  
Iron Oxide Nanoparticles ◽  
Cell Tracking ◽  
Superparamagnetic Iron Oxide ◽  
Superparamagnetic Iron Oxide Nanoparticles ◽  
Cell Populations ◽  
Oxide Nanoparticles ◽  
Signal Loss

Determination of the dynamics of specific cell populations in vivo is essential for the development of cell-based therapies. For cell tracking by magnetic resonance imaging (MRI), cells need to internalize, or be surface labeled with a MRI contrast agent, such as superparamagnetic iron oxide nanoparticles (SPIOs): SPIOs give rise to signal loss by gradient-echo and T2-weighted MRI techniques. In this study, cancer cells were chemically tagged with biotin and then magnetically labeled with anti-biotin SPIOs. No significant detrimental effects on cell viability or death were observed following cell biotinylation. SPIO-labeled cells exhibited signal loss compared to non-SPIO-labeled cells by MRI in vitro. Consistent with the in vitro MRI data, signal attenuation was observed in vivo from SPIO-labeled cells injected into the muscle of the hind legs, or implanted subcutaneously into the flanks of mice, correlating with iron detection by histochemical and X-ray fluorescence (XRF) methods. To further validate this approach, human mesenchymal stem cells (hMSCs) were also employed. Chemical biotinylation and SPIO labeling of hMSCs were confirmed by fluorescence microscopy and flow cytometry. The procedure did not affect proliferation and multipotentiality, or lead to increased cell death. The SPIO-labeled hMSCs were shown to exhibit MRI signal reduction in vitro and was detectable in an in vivo model. In this study, we demonstrate a rapid, robust, and generic methodology that may be a useful and practical adjuvant to existing methods of cell labeling for in vivo monitoring by MRI. Further, we have shown the first application of XRF to provide iron maps to validate MRI data in SPIO-labeled cell tracking studies.

scholarly journals Invasion performance-similarity found among multiple cell systems

2020 ◽  
Author(s):  
Om Prakash
Keyword(s):  
Cell Lines ◽  
Drug Repositioning ◽  
Biological Research ◽  
Multiple Systems ◽  
Biologically Relevant ◽  
Selective Targeting ◽  
Ann Models ◽  
Multiple Cell ◽  
Artificial Neural Network Ann ◽  
Network Mapping

ABSTRACTUnderstanding of inter-system behavior develops biologically relevant intuition for drug repositioning as well as other biological research. But combining all the possible genes interactions into a system, and furthermore comparisons of multiple systems are a challenge on time ground with feasible experiments. In present study, 64 cell lines from 11 different organs were compared for their invasion performance. RNA expressions of 23 genes were used to create systems artificial neural network (ANN) models. ANN models were prepared for all 64 cell lines and observed for their invasion performance through network mapping. The resulted cell line clusters bear feasible capacity to perform experiments for biologically relevant research motivations as drug repositioning and selective targeting etc.; and can be used for analysis of invasion related aspects.

scholarly journals β-catenin-mediated Wnt signal transduction proceeds through an endocytosis-independent mechanism

2020 ◽  
Vol 31 (13) ◽  
pp. 1425-1436 ◽  
Author(s):  
Ellen Youngsoo Rim ◽  
Leigh Katherine Kinney ◽  
Roeland Nusse
Keyword(s):  
Signal Transduction ◽  
Real Time ◽  
Cell Lines ◽  
Wnt Pathway ◽  
Receptor Endocytosis ◽  
Pathway Activation ◽  
Independent Mechanism ◽  
Multiple Cell ◽  
Wnt Signal ◽  
Time Assessment

A novel tool for quantitative, real-time assessment of Wnt pathway activation was combined with genetic disruption of endocytosis to determine whether receptor endocytosis is required for Wnt signal transduction. Our results in multiple cell lines support that clathrin- or caveolin-mediated endocytosis is dispensable for Wnt signal transduction.

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