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Cytosolic delivery of nanolabels prevents their asymmetric inheritance and enables extended quantitative in vivo cell imaging

Ranhua Xiong (UGent) , Freya Joris (UGent) , Sayuan Liang, Riet De Rycke (UGent) , Saskia Lippens (UGent) , Jo Demeester (UGent) , Andre Skirtach (UGent) , Koen Raemdonck (UGent) , Uwe Himmelreich, Stefaan De Smedt (UGent) , et al.
(2016) NANO LETTERS. 16(10). p.5975-5986
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Center for nano- and biophotonics (NB-Photonics)
Abstract
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 labeling 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 vapor 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 iihotoporation 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 labeling by photoporation resulted in a marked improvement in long-term cell visibility in vivo where an insulin producing cell line (INS-1E cell line) labeled with fluorescent dextrans could be tracked for up to two months in Swiss nude mice compared to 2-Weeks for cells labeled by endocytosis.
Keywords
Cell labeling cytosolic delivery, nanopaiticle inheritance, vapor nanobubble photoporation, in vivo cell tracking, quantum dots, NEURAL STEM-CELLS, QUANTUM DOTS, REPORTER GENE, LIVE CELLS, INTRACELLULAR DELIVERY, PLASMONIC NANOBUBBLES, ENDOSOMAL ESCAPE, CONTRAST AGENTS, TRACKING, DIVISION

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Citation

Please use this url to cite or link to this publication:

Chicago
Xiong, Ranhua, Freya Joris, Sayuan Liang, Riet De Rycke, Saskia Lippens, Jo Demeester, Andre Skirtach, et al. 2016. “Cytosolic Delivery of Nanolabels Prevents Their Asymmetric Inheritance and Enables Extended Quantitative in Vivo Cell Imaging.” Nano Letters 16 (10): 5975–5986.
APA
Xiong, R., Joris, F., Liang, S., De Rycke, R., Lippens, S., Demeester, J., Skirtach, A., et al. (2016). Cytosolic delivery of nanolabels prevents their asymmetric inheritance and enables extended quantitative in vivo cell imaging. NANO LETTERS, 16(10), 5975–5986.
Vancouver
1.
Xiong R, Joris F, Liang S, De Rycke R, Lippens S, Demeester J, et al. Cytosolic delivery of nanolabels prevents their asymmetric inheritance and enables extended quantitative in vivo cell imaging. NANO LETTERS. 2016;16(10):5975–86.
MLA
Xiong, Ranhua, Freya Joris, Sayuan Liang, et al. “Cytosolic Delivery of Nanolabels Prevents Their Asymmetric Inheritance and Enables Extended Quantitative in Vivo Cell Imaging.” NANO LETTERS 16.10 (2016): 5975–5986. Print.
@article{8522072,
  abstract     = {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 labeling 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 vapor 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 iihotoporation 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 labeling by photoporation resulted in a marked improvement in long-term cell visibility in vivo where an insulin producing cell line (INS-1E cell line) labeled with fluorescent dextrans could be tracked for up to two months in Swiss nude mice compared to 2-Weeks for cells labeled by endocytosis.},
  author       = {Xiong, Ranhua and Joris, Freya and Liang, Sayuan and De Rycke, Riet and Lippens, Saskia and Demeester, Jo and Skirtach, Andre and Raemdonck, Koen and Himmelreich, Uwe and De Smedt, Stefaan and Braeckmans, Kevin},
  issn         = {1530-6984},
  journal      = {NANO LETTERS},
  keyword      = {Cell labeling cytosolic delivery,nanopaiticle inheritance,vapor nanobubble photoporation,in vivo cell tracking,quantum dots,NEURAL STEM-CELLS,QUANTUM DOTS,REPORTER GENE,LIVE CELLS,INTRACELLULAR DELIVERY,PLASMONIC NANOBUBBLES,ENDOSOMAL ESCAPE,CONTRAST AGENTS,TRACKING,DIVISION},
  language     = {eng},
  number       = {10},
  pages        = {5975--5986},
  title        = {Cytosolic delivery of nanolabels prevents their asymmetric inheritance and enables extended quantitative in vivo cell imaging},
  url          = {http://dx.doi.org/10.1021/acs.nanolett.6b01411},
  volume       = {16},
  year         = {2016},
}

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