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Characterisation and propagation of tumor initiating cells derived from colorectal liver metastases: trials, tribulations and a cautionary note.

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journal contribution
posted on 20.04.2015, 13:19 by Mark I. James, Lynne M. Howells, Ankur Karmokar, Jennifer A. Higgins, Peter Greaves, Hong Cai, Ashley Dennison, Matthew Metcalfe, Giuseppe Garcea, David M. Lloyd, D. P. Berry, William P. Steward, Karen Brown
Tumor initiating cells (TIC) are increasingly being put forward as a potential target for intervention within colorectal cancer. Whilst characterisation and outgrowth of these cells has been extensively undertaken in primary colorectal cancers, few data are available describing characteristics within the metastatic setting. Tissue was obtained from patients undergoing surgical resection for colorectal liver metastases, and processed into single cell suspension for assessment. Tumor initiating cells from liver metastases were characterised using combinations of EPCAM, Aldehyde dehydrogenase activity, CD133 and CD26. CD133 expression was significantly lower in patients who had received chemotherapy, but this was accounted for by a decrease observed in the male patient cohort only. ALDHhigh populations were rare (0.4 and 0.3% for EPCAM+/ALDHhigh/CD133- and EPCAM+/ALDHhigh/CD133+ populations respectively) and below the limits of detection in 28% of samples. Spheroid outgrowth of metastatic tumor cells across all samples could not be readily achieved using standard spheroid-formation techniques, thus requiring further method validation to reliably propagate cells from the majority of tissues. Spheroid formation was not enhanced using additional growth factors or fibroblast co-culture, but once cells were passaged through NOD-SCID mice, spheroid formation was observed in 82% samples, accompanied by a significant increase in CD26. Order of spheroid forming ability was ALDHhigh>CD133>CD26. Samples sorted by these markers each had the ability to reform ALDHhigh, CD133 and CD26 positive populations to a similar extent, suggestive of a high degree of plasticity for each population. Ex vivo TIC models are increasingly being utilised to assess efficacy of therapeutic interventions. It is therefore essential that such investigations use well-characterised models that are able to sustain TIC populations across a large patient cohort in order that the inherent heterogeneity observed in cancer populations is maintained.

Funding

This work was supported by a studentship from Hope Against Cancer (http://www.hfcr.org/), Leicester, UK (KB/LMH), by Cancer Research UK (http://www.cancerresearchuk.org/)on a programme grant [C325/A13101] (KB/WPS) and by Cancer Research UK in conjunction with the UK Department of Health on an Experimental Cancer Medicine Centre grant [C325/A15575] (WPS/KB).

History

Citation

PLoS One 10(2): e0117776

Author affiliation

/Organisation/COLLEGE OF MEDICINE, BIOLOGICAL SCIENCES AND PSYCHOLOGY/School of Medicine/Department of Cancer Studies and Molecular Medicine

Version

VoR (Version of Record)

Published in

PLoS One 10(2): e0117776

Publisher

Public Library of Science

issn

1932-6203

eissn

1932-6203

Copyright date

2015

Available date

20/04/2015

Publisher version

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0117776

Language

en