Abstract
Antiangiogenic therapy for cancer is a strategy targeted at tumour vasculature, often in combination with conventional cytotoxicity treatments. Animal testing is still the most common method used for evaluating the efficacy of new drugs but tissue-engineered in vitro models are becoming more acceptable for replacing and reducing the use of animals in anti-cancer drug screening. In this study, a 3D co-culture model of human endothelial cells and ovarian cancer cells was developed. This model has the potential to mimic the interactions between endothelial cells and ovarian cancer cells. The feasibility of applying this model in drug testing was explored here. The complex morphology of the co-culture system, which features development of both endothelial tubule-like structures and tumour structures, was analysed quantitatively by an image analysis method. The co-culture morphology integrity was maintained for 10 days and the potential of the model for anti-cancer drug testing was evaluated using Paclitaxel and Cisplatin, two common anti-tumour drugs with different mechanisms of action. Both traditional cell viability assays and quantitative morphological analyses were applied in the drug testing. Cisplatin proved a good example showing the advantages of morphological analysis of the co-culture model when compared with mono-culture of endothelial cells, which did not reveal an inhibitory effect of Cisplatin on the tubule-like endothelial structures. Thus, the tubule areas of the co-culture reflected the anti-angiogenesis potential of Cisplatin. In summary, in vitro cancer models can be developed using a tissue engineering approach to more closely mimic the characteristics of tumours in vivo. Combined with the image analysis technique, this developed 3D co-culture angiogenesis model will provide more reproducible and reliably quantified results and reveal further information of the drug’s effects on both tumour cell growth and tumour angiogenesis.
Highlights
The formation of new capillary networks formed by vascular endothelial cells, is crucial in tumour development for the provision of nutrients and oxygen to a growing tumour [1,2]
In sandwich co-culture, a combination of tubules and spheroids was observed, as shown in Fig 2(C), and these structures were maintained and kept growing for 10 days. This relatively long culture time is in contrast to the fact that the tubule network formed by HUVECs cultured alone in the Matrigel sandwich showed degradation at day 2 and there were no signs of living cells at day 10 (Fig 2(A))
As the tubule structures formed by HUVECs in mono-culture already degraded at day 10 (Fig 2(A)), we had to question whether or not the tubule network was mainly formed by OVCAR8 only at day 10
Summary
The formation of new capillary networks formed by vascular endothelial cells, is crucial in tumour development for the provision of nutrients and oxygen to a growing tumour [1,2]. The high levels of laminin and growth factors within Matrigel promote vascular endothelial cells to form tubule-like structures [5] This is a short-term assay, because the tubule structures formed by HUVECs degrade after 48 hours [6] whereas in the in vivo environment the angiogenesis process would take place in a few days. The aim of this study, was to develop a 3D in vitro co-culture model for cancer angiogenesis studies to test drugs with anti-angiogenesis potential This co-culture model features both a vascular network and tumour structures in a 3D environment with an imaging technique (i.e. image segmentation utilised to provide quantitative results), without the need for further fluorescence labelling
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