Long-term in vivo monitoring of mouse and human hematopoietic stem cell engraftment with a human positron emission tomography reporter gene

McCracken, Melissa N.; Gschweng, Eric H.; Nair-Gill, Evan; McLaughlin, Jami; Cooper, Aaron; Riedinger, Mireille; Cheng, Donghui; Nosala, Christopher; Kohn, Donald B.; Witte, Owen N.

doi:10.1073/pnas.1221840110

Cited by 40 publications

(47 citation statements)

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“…One is to transfect the cells with a reporter gene 2,7 and follow-up on their proliferation through optical imaging, positron emission tomography or magnetic resonance imaging [8][9][10] . This strategy, however, requires a rather time-consuming transfection procedure that may lead to undesired phenotypical alterations and abnormalities in the transplanted cells in comparison to the native cells [11][12][13] .…”

Section: Introductionmentioning

confidence: 99%

Cytosolic Delivery of Nanolabels Prevents Their Asymmetric Inheritance and Enables Extended Quantitative in Vivo Cell Imaging

et al. 2016

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Long-term in vivo imaging of cells is crucial for the understanding of cellular fate in biological processes in cancer research, immunology or in cell-based therapies such as beta cell transplantation in type I diabetes or stem cell therapy. Traditionally, cell labelling with the desired contrast agent occurs ex vivo via spontaneous endocytosis, which is a variable and slow process that requires optimization for each particular label-cell type combination. Following endocytic uptake, the contrast agents mostly remain entrapped in the endolysosomal compartment, which leads to signal instability, cytotoxicity and asymmetric inheritance of the labels upon cell division. Here, we demonstrate that these disadvantages can be circumvented by delivering contrast agents directly into the cytoplasm via vapour nanobubble photoporation. Compared to classic endocytic uptake, photoporation resulted in 50 and 3 times higher loading of fluorescent dextrans and quantum dots, respectively, with improved signal stability and reduced cytotoxicity. Most interestingly, cytosolic delivery by photoporation prevented asymmetric inheritance of labels by daughter cells over subsequent cell generations. Instead, unequal inheritance of endocytosed labels resulted in a dramatic increase in polydispersity of the amount of labels per cell with each cell division, hindering accurate quantification of cell numbers in vivo over time. The combined benefits of cell labelling by photoporation resulted in a marked improvement in long-term cell visibility in vivo where an insulin producing cell line (INS-1E cell line) labelled with fluorescent dextrans could be tracked for up to two months in Swiss Nude mice compared to two weeks for cells labelled by endocytosis.

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Section: Introductionmentioning

confidence: 99%

Cytosolic Delivery of Nanolabels Prevents Their Asymmetric Inheritance and Enables Extended Quantitative in Vivo Cell Imaging

et al. 2016

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“…An adaptation of nonrestrictive linear amplification-mediated PCR (nrLAM-PCR) was used to produce integration site sequencing libraries for Illumina sequencing. 5,[15][16][17][18][19][20][21] …”

Section: Integration Site Sequencingmentioning

confidence: 99%

Cytoreductive conditioning intensity predicts clonal diversity in ADA-SCID retroviral gene therapy patients

Cooper

Lill²,

Shaw³

et al. 2017

Blood

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“…126 The technical advantages and disadvantages of imaging machinery such as fluorescence imaging, magnetic resonance imaging, positron emission tomography, single photon emission computed tomography, and bioluminescence imaging are well described elsewhere. 127 …”

Section: Technology Advancement For Integrative Stem Cell Analysesmentioning

confidence: 99%

Technology Advancement for Integrative Stem Cell Analyses

Jeong

Choi

Lee

2014

Tissue Engineering Part B: Reviews

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Scientists have endeavored to use stem cells for a variety of applications ranging from basic science research to translational medicine. Population-based characterization of such stem cells, while providing an important foundation to further development, often disregard the heterogeneity inherent among individual constituents within a given population. The population-based analysis and characterization of stem cells and the problems associated with such a blanket approach only underscore the need for the development of new analytical technology. In this article, we review current stem cell analytical technologies, along with the advantages and disadvantages of each, followed by applications of these technologies in the field of stem cells. Furthermore, while recent advances in micro/nano technology have led to a growth in the stem cell analytical field, underlying architectural concepts allow only for a vertical analytical approach, in which different desirable parameters are obtained from multiple individual experiments and there are many technical challenges that limit vertically integrated analytical tools. Therefore, we propose-by introducing a concept of vertical and horizontal approach-that there is the need of adequate methods to the integration of information, such that multiple descriptive parameters from a stem cell can be obtained from a single experiment.

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Long-term in vivo monitoring of mouse and human hematopoietic stem cell engraftment with a human positron emission tomography reporter gene

Cited by 40 publications

References 45 publications

Cytosolic Delivery of Nanolabels Prevents Their Asymmetric Inheritance and Enables Extended Quantitative in Vivo Cell Imaging

Cytosolic Delivery of Nanolabels Prevents Their Asymmetric Inheritance and Enables Extended Quantitative in Vivo Cell Imaging

Cytoreductive conditioning intensity predicts clonal diversity in ADA-SCID retroviral gene therapy patients

Technology Advancement for Integrative Stem Cell Analyses

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