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Mark I. James
Mark I.
James
Lynne M. Howells
Lynne M.
Howells
Ankur Karmokar
Ankur
Karmokar
Jennifer A. Higgins
Jennifer A.
Higgins
Peter Greaves
Peter
Greaves
Hong Cai
Hong
Cai
Ashley Dennison
Ashley
Dennison
Matthew Metcalfe
Matthew
Metcalfe
Giuseppe Garcea
Giuseppe
Garcea
David M. Lloyd
David M.
Lloyd
D. P. Berry
D. P.
Berry
William P. Steward
William P.
Steward
Karen Brown
Karen
Brown
Characterisation and propagation of tumor initiating cells derived from colorectal liver metastases: trials, tribulations and a cautionary note.
University of Leicester
2015
Chemotherapy
Colorectal cancer
Cell differentiation
Flow cytometry
Hepatic resection
Tumor stem cells
Growth factors
Proteomes
2015-04-20 13:19:41
Journal contribution
https://figshare.le.ac.uk/articles/journal_contribution/Characterisation_and_propagation_of_tumor_initiating_cells_derived_from_colorectal_liver_metastases_trials_tribulations_and_a_cautionary_note_/10164266
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.