@article{b53fc876c92740b7abebc7028d2a5152,
title = "In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy",
abstract = "Quantification of cell-cycle state at a single-cell level is essential to understand fundamental three-dimensional (3D) biological processes such as tissue development and cancer. Analysis of 3D in vivo images, however, is very challenging. Today's best practice, manual annotation of select image events, generates arbitrarily sampled data distributions, which are unsuitable for reliable mechanistic inferences. Here, we present an integrated workflow for quantitative in vivo cell-cycle profiling. It combines image analysis and machine learning methods for automated 3D segmentation and cell-cycle state identification of individual cell-nuclei with widely varying morphologies embedded in complex tumor environments. We applied our workflow to quantify cell-cycle effects of three antimitotic cancer drugs over 8 d in HT-1080 fibrosarcoma xenografts in living mice using a data set of 38,000 cells and compared the induced phenotypes. In contrast to results with 2D culture, observed mitotic arrest was relatively low, suggesting involvement of additional mechanisms in their antitumor effect in vivo.",
author = "Chittajallu, {Deepak R.} and Stefan Florian and Kohler, {Rainer H.} and Yoshiko Iwamoto and Orth, {James D.} and Ralph Weissleder and Gaudenz Danuser and Mitchison, {Timothy J.}",
note = "Funding Information: S.F. was supported by a research fellowship (FL 820/1-1) from the DFG (Deutsche Forschungsgemeinschaft). This project was supported by US National Institutes of Health grants R01-CA164448, S10RR0266360 and PO1-CA139980. We thank P. Choi for helpful discussions. We are grateful for the support of The Nikon Imaging Center at Harvard Medical School. We thank K. Krukenberg (Harvard Medical School) for the MCF7 and T47D cell lines stably expressing H2B-GFP; P. Keller (Howard Hughes Medical Institute, Janelia Research Campus) for Drosophila, mouse and zebrafish data sets; M. Sebas for surgical implantation of the DSCs; and J. Moore for assistance with flow cytometry. The FUCCI viral particles were a kind gift from P. Jorgensen, A. Tzur and M. Chung (Harvard Medical School) generated with vectors kindly provided by the laboratory of A. Miyawaki (RIKEN Brain Science Institute). The H2B-CFP construct was obtained from A. Loewer (Max Delbrueck Center). Publisher Copyright: {\textcopyright} 2015 Nature America, Inc. All rights reserved.",
year = "2015",
month = may,
day = "28",
doi = "10.1038/nmeth.3363",
language = "English (US)",
volume = "12",
pages = "577--585",
journal = "Nature Methods",
issn = "1548-7091",
publisher = "Public Library of Science",
number = "6",
}