In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy

Deepak R. Chittajallu, Stefan Florian, Rainer H. Kohler, Yoshiko Iwamoto, James D. Orth, Ralph Weissleder, Gaudenz Danuser, Timothy J. Mitchison

Research output: Contribution to journalArticle

48 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)577-585
Number of pages9
JournalNature Methods
Volume12
Issue number6
DOIs
StatePublished - May 28 2015

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Heterografts
Tumors
Cell Cycle
Cells
Workflow
Neoplasms
Antimitotic Agents
Biological Phenomena
Fibrosarcoma
Cell Nucleus
Practice Guidelines
Image analysis
Learning systems
Tissue
Phenotype
Intravital Microscopy
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology

Cite this

Chittajallu, D. R., Florian, S., Kohler, R. H., Iwamoto, Y., Orth, J. D., Weissleder, R., ... Mitchison, T. J. (2015). In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy. Nature Methods, 12(6), 577-585. https://doi.org/10.1038/nmeth.3363

In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy. / Chittajallu, Deepak R.; Florian, Stefan; Kohler, Rainer H.; Iwamoto, Yoshiko; Orth, James D.; Weissleder, Ralph; Danuser, Gaudenz; Mitchison, Timothy J.

In: Nature Methods, Vol. 12, No. 6, 28.05.2015, p. 577-585.

Research output: Contribution to journalArticle

Chittajallu, DR, Florian, S, Kohler, RH, Iwamoto, Y, Orth, JD, Weissleder, R, Danuser, G & Mitchison, TJ 2015, 'In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy', Nature Methods, vol. 12, no. 6, pp. 577-585. https://doi.org/10.1038/nmeth.3363
Chittajallu DR, Florian S, Kohler RH, Iwamoto Y, Orth JD, Weissleder R et al. In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy. Nature Methods. 2015 May 28;12(6):577-585. https://doi.org/10.1038/nmeth.3363
Chittajallu, Deepak R. ; Florian, Stefan ; Kohler, Rainer H. ; Iwamoto, Yoshiko ; Orth, James D. ; Weissleder, Ralph ; Danuser, Gaudenz ; Mitchison, Timothy J. / In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy. In: Nature Methods. 2015 ; Vol. 12, No. 6. pp. 577-585.
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