Lina Chakrabarti, Christopher Groves and Michael Lee on the Challenges with Therapeutic Cell Sorting
Two weeks ago, the Cellix team travelled to Prague to participate in Cyto 2018, a conference discussing the cutting edge of flow cytometry research. In the second of a 4 part series, here's our key take-aways from the conference.
As I mentioned in a previous post, one of the most interesting points differentiating Cyto 2018 from previous years was the focus on flow cytometry use in human gene therapy.
The importance placed on the topic this year was shown in the choice of lectures - this year's Cyto Innovation keynote talk was by Dr. Serena Scala from the San Rafaelle-Telethon Institute for Gene Therapy, on the subject of 'Genetic Engineering of Haematopoiesis to Treat Inherited Diseases'. In it, she discussed some of her latest work with novel methods for characterisation of haematopoietic disorders and patient monitoring after transplant or gene therapy.
Challenges in Therapeutic Cell Sorting: Christopher Groves
Christopher J. Groves, senior R&D manager at MedImmune, moderated an extremely interesting Therapeutic Cell Sorting Workshop. Chris had earlier given a really interesting talk focussed on automation of sample prep, process assays and the resulting positive effect had on cell viability at the Miltenyi workshops, so I decided to sit in on the therapeutic cell sorting workshop out of interest. From my point of view, this was one of the ultimate highlights of the conference.
Christopher gave an overview of a survey results on requirements for therapeutic cell sorting. This provided interactive engagement with the audience on some of the challenges facing the development of tools and techniques in this field. Particularly, the discussions on the types of fluidics used in cell sorter systems, ensuring sterility, and the disadvantages of jet-in-air sorters typify the difficulties I've noticed researchers in the field facing.
The Downsides of Dyes
One other point of interest from the workshop was the discussion on the effect of GFP, dyes & fluorescent strains on cell function. This is something which has been historically overlooked but is gaining attention - particularly as the therapeutic aspect of flow cytometry becomes more prominent, the potential inflammatory effects of stained cells on re-insertion becomes more important.
Personally, I think the decreasing cost of tools like ChIP (to measure protein-DNA interactions) and rt-qPCR (to measure differences in gene expression) will lead to some interesting small studies on cell function before and after the addition of these plasmids/ dyes/ stains. I see this emerging information continuing to drive the trend of interest in label-free techniques. Given the ubiquity of these methods, this is definitely a space to watch!
Cell-sorting in Industry Challenges: Lina Chakrabarti
Lina Chakrabarti from MedImmune gave a fascinating talk entitled 'Flow Cytometry: An Indispensable Tool for Isolating High-Producing Clonal Cell Lines for Biopharmaceutical Manufacture'. In it, she discussed the development of cell lines for bio therapeutic protein production, and some of the challenges associated with stable cell generation and ensuring monoclonality.
In the issues Lina discussed I see a reflection of a common industry need: the requirement for high-throughput screening of live single cells for markers predictive of monoclonality, and the pressing need for methods to enhance the selection efficiency of rare high-yielding monoclonal production cell lines.
Overall, the take home from Cyto 2018 on therapeutic cell sorting is that the nascent application of flow in this field could make a real impact on the efficacy of treatments, but currently faces numerous challenges.
Improving Cell-Sorting from the Perspective of Tool Companies: Michael Lee
Michael Lee is the flow cytometry core Director at the University of California San Francisco. He gave a talk on what is required from tool providers to improve products for sorting transfected cells. He mentioned that for their work they were given FDA approval for Phase I and II, but that this was only on compassionate grounds - the FDA mentioned that for Phase III, the system they use must be closed loop.
Michael also discussed single cell sorting efficiency and cell outgrowth, issues around sterility, biomanufacturing and use of the BD FACS Aria for cell sorting.
Owl BioMedical work on advancing MEMS as a sterile cell-sorting platform, and their consultant Jack Dunne gave some valuable comments at this workshop.
For more info, read our other Cyto 2018 posts on: