Performing a cell killing assay without using radioactive materials

Share this:

Have you ever thought of performing a cell killing assay without using radioactive materials?

Phenomics Australia node at the VCFG recently teamed up with Dr Kevin Sek to develop an application using their Incucyte SX5 Live Cell Analysis instrument.

Dr Sek tracked live tumour (adherent and non-adherent) cell growth over 48 hours in the presence or absence of CAR T cells. He was able to show that CAR T cells were effectively killing tumour cells over time. He captured images and videos of the whole process in 384-well plate format, allowing him to test several conditions in a single experiment. In comparison, if he performed the gold standard chromium killing assay, in a 96-well format, he would use substantially more cells, would be limited in the number of test conditions and would only have a single end point assay result.

This is such a great advance!

Dr Kevin Sek (R), Postdoctoral Researcher at the Peter MacCallum Cancer Centre and Dr Henry Beetham (L), Research Officer at the VCFG, Peter MacCallum Cancer Centre.
Dr Kevin Sek, loading the Incucyte SX5 Live Cell instrument at the Victorian Centre For Functional Genomics (VCFG) at Peter MacCallum Cancer Centre.
Human MCF7 adherent breast tumour cells where incubated alone or in the presence of CAR T cells in the Incucyte. Images were taken every 4 hours over 48 hours. MCF7 cells were stained red with NucLight and the number of live red stained tumour cells were counted over time. Cell death was measured through Caspase 3/7 green flourescence.

‘ Through our collaboration with Prescient Therapeutics, we wanted to determine the anti-tumour capabilities of Prescient’s new universal OmniCAR T cell platform. I teamed up with the VCFG staff who were critical in optimizing and setting up real-time, coculture experiments on the Incucyte. Using a small number of CAR T cells and tumour targets, I was able to obtain high quality videos and images, quantify tumour cell killing and T cell proliferation, and also determine multiple antigen targeting of the Universal OmniCAR T cells in a mixed tumour killing assay.”

Dr Kevin Sek
Dr Kevin Sek, Postdoctoral Researcher at the Peter MacCallum Cancer Centre.

The IncuCyte SX5, at the Victorian Centre for Functional Genomics (VCFG), is a live-cell imaging microscope that is housed inside a cell culture incubator. It allows researchers to image up to six 384-well plates in phase, green and red channels every 2 hours, in parallel! Its user friendly software can quantify cell number, size, shape and fluorescence intensity at the single-cell level over time. 

Phenomics Australia Feedback Form

Phenomics Australia (formerly the Australian Phenomics Network, APN) offers consolidated infrastructure and expertise supporting genomic medicine and biomedical research from discovery through into early clinical development and evaluation.

Phenomics Australia is a founding capability enabled by the National Collaborative Research Infrastructure Strategy (NCRIS). We would love to hear from you!

Please feel free to provide any ideas or general comments you might have so that we can further improve how Phenomics Australia can support you.

Phenomics Australia PHENOMENA News Contribution

Have you recently had a publication accepted? Do you want to tell us about a grant you have received? Or maybe you have just been recently in the news? You can complete this form to tell us about your latest research developments.

Information provided will be used to inform press releases, website stories, social media content and more. This form is being used to capture everything that is happening at Phenomics Australia, which can then be used as part of our outreach and communications.

Once you have completed the form, the Communications and Outreach Coordinator will then be in contact with you.

Thanks.

Make an Enquiry

_Get in touch to discuss how we can help with your research.