Advertisement

Cell Migration pp 163-173 | Cite as

Cell Migration in Tissues: Explant Culture and Live Imaging

  • Ralitza Staneva
  • Jorge Barbazan
  • Anthony Simon
  • Danijela Matic VignjevicEmail author
  • Denis Krndija
Part of the Methods in Molecular Biology book series (MIMB, volume 1749)

Abstract

Cell migration is a process that ensures correct cell localization and function in development and homeostasis. In disease such as cancer, cells acquire an upregulated migratory capacity that leads to their dissemination throughout the body. Live imaging of cell migration allows for better understanding of cell behaviors in development, adult tissue homeostasis and disease. We have optimized live imaging procedures to track cell migration in adult murine tissue explants derived from: (1) healthy gut; (2) primary intestinal carcinoma; and (3) the liver, a common metastatic site. To track epithelial cell migration in the gut, we generated an inducible fluorescent reporter mouse, enabling us to visualize and track individual cells in unperturbed gut epithelium. To image intratumoral cancer cells, we use a spontaneous intestinal cancer model based on the activation of Notch1 and deletion of p53 in the mouse intestinal epithelium, which gives rise to aggressive carcinoma. Interaction of cancer cells with a metastatic niche, the mouse liver, is addressed using a liver colonization model. In summary, we describe a method for long-term 3D imaging of tissue explants by two-photon excitation microscopy. Explant culturing and imaging can help understand dynamic behavior of cells in homeostasis and disease, and would be applicable to various tissues.

Key words

Tissue explant Cell migration Two-photon imaging Ex vivo culture Gut Tumor Liver 

Notes

Acknowledgments

We would like to thank Basile Gurchenkov and Fatima El Marjou for assistance with microscopes and mice. The authors greatly acknowledge the Cell and Tissue Imaging (PICT-IBiSA), Institut Curie, member of the French National Research Infrastructure France-BioImaging (ANR10-INBS-04). This work was supported by the Fondation pour la Recherche Médicale (FRM N° DGE20111123020), the Canceropole-IdF (nis2012-2-EML-04-IC-1), INCa (Cancer National Institute, n° 2011-1-LABEL-IC-4). The authors would like to acknowledge the Cell and Tissue Imaging Platform - PICT-IBiSA (member of France–Bioimaging, ANR-10-INBS-04) of the Genetics and Developmental Biology Department (UMR3215/U934) of Institut Curie for help with image analysis.

This work is funded by Institut Thématique Multi-organismes Cancer–Plan Cancer 2014–2019 and Ecole Doctorale Frontières du Vivant (FdV)–Programme Bettencourt (RS), Marie Curie Individual Fellowship (FiBRO) (JB), and ERC starting grant (DMV).

References

  1. 1.
    Lambert AW, Pattabiraman DR, Weinberg RA (2017) Emerging biological principles of metastasis. Cell 168:670–691.  https://doi.org/10.1016/j.cell.2016.11.037 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Chambers AF, Groom AC, MacDonald IC (2002) Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer 2:563–572.  https://doi.org/10.1038/nrc865 CrossRefPubMedGoogle Scholar
  3. 3.
    Valastyan S, Weinberg RA (2011) Tumor metastasis: molecular insights and evolving paradigms. Cell 147:275–292.  https://doi.org/10.1016/j.cell.2011.09.024 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Friedl P, Wolf K (2010) Plasticity of cell migration: a multiscale tuning model. J Cell Biol 188:11–19.  https://doi.org/10.1083/jcb.200909003 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Clark AG, Vignjevic DM (2015) Modes of cancer cell invasion and the role of the microenvironment. Curr Opin Cell Biol 36:13–22.  https://doi.org/10.1016/j.ceb.2015.06.004 CrossRefPubMedGoogle Scholar
  6. 6.
    Kedrin D, Gligorijevic B, Wyckoff J, Verkhusha VV, Condeelis J, Segall JE, van Rheenen J (2008) Intravital imaging of metastatic behavior through a mammary imaging window. Nat Methods 5:1019–1021.  https://doi.org/10.1038/nmeth.1269 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Alexander S, Koehl GE, Hirschberg M, Geissler EK, Friedl P (2008) Dynamic imaging of cancer growth and invasion: a modified skin-fold chamber model. Histochem Cell Biol 130:1147–1154.  https://doi.org/10.1007/s00418-008-0529-1 CrossRefPubMedGoogle Scholar
  8. 8.
    Muzumdar MD, Tasic B, Miyamichi K, Li L, Luo L (2007) A global double-fluorescent Cre reporter mouse. Genesis 45:593–605.  https://doi.org/10.1002/dvg.20335 CrossRefPubMedGoogle Scholar
  9. 9.
    El Marjou F, Janssen KP, Chang BHJ, Li M, Hindie V, Chan L, Louvard D, Chambon P, Metzger D, Robine S (2004) Tissue-specific and inducible Cre-mediated recombination in the gut epithelium. Genesis 39:186–193.  https://doi.org/10.1002/gene.20042 CrossRefPubMedGoogle Scholar
  10. 10.
    Chanrion M, Kuperstein I, Barrière C, El Marjou F, Cohen D, Vignjevic D, Stimmer L, Paul-Gilloteaux P, Bièche I, Tavares SDR, Boccia G-F, Cacheux W, Meseure D, Fre S, Martignetti L, Legoix-Né P, Girard E, Fetler L, Barillot E, Louvard D, Zinovyev A, Robine S (2014) Concomitant Notch activation and p53 deletion trigger epithelial-to-mesenchymal transition and metastasis in mouse gut. Nat Commun 5:5005.  https://doi.org/10.1038/ncomms6005 CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Barbazán J, Alonso-Alconada L, Elkhatib N, Geraldo S, Gurchenkov V, Glentis A, Van Niel G, Palmulli R, Fernández B, Viaño P, Garcia-Caballero T, López-López R, Abal M, Vignjevic DM, Barbazan J (2017) Liver metastasis is facilitated by the adherence of circulating tumor cells to vascular fibronectin deposits. Cancer Res 77(13):3431–3441.  https://doi.org/10.1158/0008-5472.CAN-16-1917 CrossRefPubMedGoogle Scholar
  12. 12.
    Van Cutsem E, Nordlinger B, Adam R, Köhne C-H, Pozzo C, Poston G, Ychou M, Rougier P (2006) Towards a pan-European consensus on the treatment of patients with colorectal liver metastases. Eur J Cancer 42:2212–2221.  https://doi.org/10.1016/j.ejca.2006.04.012 CrossRefPubMedGoogle Scholar
  13. 13.
    Parslow A, Cardona A, Bryson-Richardson RJ (2014) Sample drift correction following 4D confocal time-lapse imaging. J Vis Exp.  https://doi.org/10.3791/51086
  14. 14.
    Meijering E, Dzyubachyk O, Smal I (2012) Methods for cell and particle tracking. Methods Enzymol 504:183–200.  https://doi.org/10.1016/B978-0-12-391857-4.00009-4 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Ralitza Staneva
    • 1
    • 2
  • Jorge Barbazan
    • 1
  • Anthony Simon
    • 1
  • Danijela Matic Vignjevic
    • 1
    Email author
  • Denis Krndija
    • 1
  1. 1.Institut CuriePSL Research University, CNRS, UMR 144ParisFrance
  2. 2.University Paris DescartesParisFrance

Personalised recommendations