Biotechnology and Biological Sciences Doctoral Training Programme
   
   
  

Investigation of Metastatic Cancers using a Perfusion Culture Bioreactor

 

Lab rotation project description

Substrate preference of breast cancer cells. Metastasising cancer cells   from specific primary origins tend to recolonise particular secondary sites, for example breast cancer cells most commonly disseminate to the lymph nodes, lungs  and bone. Their choice of destination depends largely on the composition of the extracellular matrices present at the secondary location. The aim of this project is to optimise decellularisation protocols for various bovine (or porcine) tissues to enable breast cancer colonisation and harvesting to be achieved with sufficient yield for downstream applications, including clonogenic survival and analysis of genetic stability. 

Fact file

Research theme

IBB

Location

Life Sciences

Rotation

LR2 and LR3

Contact

2nd supervisor


BBSRC Doctoral Training Partnerships
 

Linked PhD Project Outline

In most   cancer patients, primary tumours can be surgically resected and/or   irradiated, thus they are only life-threatening in specific locations, such   as the brain and pancreas. However, once a cancer spreads to other tissues it   almost invariably becomes lethal: upon dissemination from the primary site to   form secondaries, the only option is systemic chemotherapy, which can prolong   life but does not provide a cure. In the lab, cell motility assays such as   the scratch test and migration through a Boyden chamber are traditionally   used as a model for cancer metastasis.  However these monolayer-based   assays are wholly inadequate for this purpose as they do not mimic the native   3D environment, the transit of cells through the blood and lymphatic systems,   or their subsequent anchoring at distal locations. The aim of this   studentship is to develop a biomimetic environment that accurately simulates   conditions encountered by metastatic cancer cells in the body, from which   samples can be taken at strategic times and locations. 

Perfusion   bioreactors are commonly used in normoxic conditions by tissue engineers to   enable high density culture of cells on 3 dimensional substrates.   Decellularised tissues of xenogenic origin offer a useful analogue for the   tissues to which metastatic tumours would be expected to spread. This project   will apply the tissue engineering and biomaterials expertise of co-I,   Alastair Campbell-Ritchie, to develop bio-mimetic micro-environments for   cancer research and the expertise of Nicola Everitt (Engineeting) in light   sheet microscopy to image tumour growth and behaviour in real time.

Work   item 1: To assess the efficacy of cancer cell   colonisation on porous 3D-scaffolds in a continuously perfused bioreactor.   Initial experiments will use highly metastatic breast cancer cells,   MDA-MB-231, expressing fluorescent reporters. Cell survival, motility and   colonisation will be assessed by fluorescent sheet microscopy in real time.   PI  Wheatley (Life Sciences, 60%),   Ritchie (Engineering, 40%).

Work   item 2: To determine cell viability during convective   transfer from primary to secondary tumours.  The bioreactor substrate   will be seeded with a specified number of cells and culture medium sampled at   intervals. The number of viable cells collected will be analysed using a   clonogenic survival  assay and morphological / biomarker   expression. 

Work   item 3: To determine the efficacy of metastasis to   distinct substrates. Within the perfusion bioreactor, the primary tumour will   be grown in one chamber and the decellularised target tissue in a   second.  ForMDA-MB-231 cells, the   target tissues will be decellularised lung (soft tissue) and cancellous bone   (hard tissue), to mimic the most common sites to which they disseminate.   Cells within the target tissue will be characterised for expression of   metastasis-biomarkers to identify tumours with the greatest metastatic   potential. 
Expected   outcome: The development of a perfusion bioreactor   optimised for investigation of cancer metastasis.
 

Biotechnology and Biological Sciences Doctoral Training Programme

The University of Nottingham
University Park
Nottingham, NG7 2RD

Tel: +44 (0) 115 8466946
Email: bbdtp@nottingham.ac.uk