Research Papers:

Short term ex-vivo expansion of circulating head and neck tumour cells

Arutha Kulasinghe, Chris Perry, Majid E. Warkiani, Tony Blick, Anthony Davies, Ken O'Byrne, Erik W. Thompson, Colleen C. Nelson, Ian Vela and Chamindie Punyadeera _

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Oncotarget. 2016; 7:60101-60109. https://doi.org/10.18632/oncotarget.11159

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Arutha Kulasinghe1,7, Chris Perry2, Majid E. Warkiani3, Tony Blick1,7, Anthony Davies4,7, Ken O'Byrne4,7, Erik W. Thompson1,7, Colleen C. Nelson5,7, Ian Vela5,6,7, Chamindie Punyadeera1,7

1The School of Biomedical Sciences, Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, QLD, Australia

2Department of Otolaryngology, Princess Alexandra Hospital, Woolloongabba, QLD, Australia

3School of Mechanical and Manufacturing Engineering, Australian Centre for NanoMedicine, University of New South Wales, Sydney, Australia

4Translational Cell Imaging Queensland, Institute of Health and Biomedical Innovation, Queensland University of Technology, QLD, Australia

5Australian Prostate Cancer Research Centre-Queensland, Institute of Health and Biomedical Innovation, Queensland University of Technology, Princess Alexandra Hospital, QLD, Australia

6Department of Urology, Princess Alexandra Hospital, Wolloongabba, QLD, Australia

7Translational Research Institute, Woolloongabba, QLD, Australia

Correspondence to:

Chamindie Punyadeera, email: [email protected]

Keywords: circulating tumour cells, head and neck cancer, ex-vivo culture, metastasis, HPV

Received: June 03, 2016     Accepted: July 20, 2016     Published: August 09, 2016


Minimally invasive techniques are required for the identification of head and neck cancer (HNC) patients who are at an increased risk of metastasis, or are not responding to therapy. An approach utilised in other solid cancers is the identification and enumeration of circulating tumour cells (CTCs) in the peripheral blood of patients. Low numbers of CTCs has been a limiting factor in the HNC field to date. Here we present a methodology to expand HNC patient derived CTCs ex-vivo. As a proof of principle study, 25 advanced stage HNC patient bloods were enriched for circulating tumour cells through negative selection and cultured in 2D and 3D culture environments under hypoxic conditions (2% O2, 5% CO2). CTCs were detected in 14/25 (56%) of patients (ranging from 1–15 CTCs/5 mL blood). Short term CTC cultures were successfully generated in 7/25 advanced stage HNC patients (5/7 of these cultures were from HPV+ patients). Blood samples from which CTC culture was successful had higher CTC counts (p = 0.0002), and were predominantly from HPV+ patients (p = 0.007). This is, to our knowledge, the first pilot study to culture HNC CTCs ex-vivo. Further studies are warranted to determine the use of short term expansion in HNC and the role of HPV in promoting culture success.

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