Customer story: Rescuing low viability cell products

In December 2020, we ran a Holiday Season Competition and a few lucky customers won a NucleoCounter® NC-202™ for a year to support their research projects. Here, we catch up with one of the winners to find out how their project is going. We speak to PhD student Anqi Li from the Hudson Institute of Medical Research, who has been developing a new process to remove non-viable cells and rescue low-viability products.

The gap between R&D and commercial-scale manufacturing of cell therapies can sometimes seem like an insurmountable chasm. Cell therapy manufacturing must be robust, cost-effective, and reproducible at the industrial scale to ensure a sufficient supply of high-quality products for patients. Automation plays a crucial role in achieving this, but commercial-scale automated manufacturing processes require significant planning and investment.

As a result, cell therapies still in clinical trials are often produced in academic settings. However, academic laboratories typically rely on time-consuming, laborious, and unreliable manual techniques in their cell therapy manufacturing processes. This makes scaling up and providing a consistent supply of therapies for patients difficult.

Bridging the gap between R&D and commercial cell manufacturing

In 2016, David James and his team from Scinogy, a company specializing in cell therapy manufacturing scale-up, recognized the need for flexible, cost-effective, small-batch manufacturing solutions that fill the gap between R&D and commercial manufacturing of advanced therapy medicinal products (ATMPs). They began developing a multi-purpose automated device for cell separation, concentration, washing, buffer exchange, and cryopreservation. Scinogy designed the device to eliminate some of the most complex and laborious manual processes in cell therapy production, therefore easing the transition from R&D to clinical manufacturing.^1,2

Scinogy developed the first prototypes of their device using beads to represent cells. But they soon realized that they needed to test it on real cells to optimize the process conditions to ensure maximum efficiency while avoiding cell damage or loss. They formed a collaboration with Associate Professor Rebecca Lim from Hudson Institute and recruited Anqi Li as a PhD student to optimize the device for use on real cells in collaboration with Thermo Fisher Scientific. Anqi has spent the last few years developing the cell washing device, which has now been successfully launched through Thermo Fisher Scientific and named the Gibco™ CTS™ Rotea™ Counterflow Centrifugation System.

Cell counting was critical in developing the Gibco™ CTS™ Rotea™

In cell therapy manufacturing, especially autologous therapies, every cell is precious. As a result, ensuring the Gibco™ CTS™ Rotea™ provided gentle processing and high live cell recovery was critically important. Anqi investigated the effects of factors including flow rate, centrifugal force, and buffer density on cell number and viability.^3,4

“When we were developing the device, cell count was critical to informing us of failure or success for every run,” says Anqi. Cell counting helped the team understand how the varying process parameters affected their cells, so that they could optimize the Gibco™ CTS™ Rotea™. But soon, manual cell counting became a bottleneck, slowing down the development process. “We were frustrated with manual counting every time,” says Anqi. “We did try other automated cell counters, but their utility came with big limitations.”

The team soon heard about the NucleoCounter® NC-202™, our automated cell counter that gives precise results in less than a minute. They didn’t hesitate to enter our competition, hoping that the NC-202™ would give them a reliable way to count their cells and improve their process development. They won, and the NC-202™ was installed in the Hudson Institute laboratory in February 2021.

Rescuing low viability cell products

The Gibco™ CTS™ Rotea™ Counterflow Centrifugation System was launched in October 2020. But the team has continued improving the system and working on ways to enhance small-scale cell therapy manufacturing.

Since winning the NC-202™, they have been working on a process to improve cell viability in ATMPs. “Cell viability is an important release criterion for cell manufacturing. If you have a batch of cells with low viability, you can’t infuse it into a patient even if you’ve achieved the target yield after expansion or collection. The batch has essentially failed,” Anqi explains.

The team hoped to make a fast and inexpensive way to rescue these low-viability products. “We developed a process that uses an additional step with the Gibco™ CTS™ Rotea™ system to remove the dead cells and improve the viability of the product,” she says.

In this study, they used the NC-202™ to measure the count and viability of their cells before and after the rescue process. “Using the NC-202™ has proven to be a good approach. Using the traditional cell counting method, we could not differentiate between the magnetic beads that we use to activate the cells and the T cells themselves,” says Anqi. The NC-202™ uses fluorescent dyes that bind to nucleic acids to detect cells, easily differentiating between cells and magnetic beads.

Their latest work was published in Cytotherapy in March this year, less than a year after the NC-202™ was installed. In the published work, they demonstrate the optimized rescue process. They report successfully improving the viability of expanded T cells from 80.7 to 94.7% and freshly isolated human amniotic epithelial cells from 79.2 to 90.3%.⁵ These are remarkable results in an industry where every cell counts.

“A reliable automated cell counter like the NC-202™ certainly made the process development a lot easier,” says Anqi.

Click here to learn more about the NucleoCounter® NC-202™.

Further reading

Journal post: All your cell counting questions, answered
More about the Gibso™ CTS™ Rotea™: Gibco CTS Rotea Counterflow Centrifugation System
Journal post: Cell counting in GMP environments
Product: NucleoCounter® NC-202™

References

1. Li, A., James, D., Lim, R. (2021), The Gibco™ CTS™ Rotea™ system story—a case study of industry-academia collaboration Gene Therapy.
2. Li, A., et al. (2021) Advances in automated cell washing and concentration Cytotherapy 23(9):774-786.
3. Li A., et al. (2020) Design of Experiment (DoE) approach to identify critical parameters in a counterflow centrifugation system Cytotherapy 22(5):S151-S152.
4. Li A., et al. (2019) Evaluating automated buffer exchange protocols using Rotea™ counterflow centrifuge Cytotherapy 21(5):S40.
5. Li A., et al. (2022) Improving cell viability using counterflow centrifugal elutriation Cytotherapy.

Gibco™ CTS™ and Rotea™ are trademarks of Thermo Fisher Scientific.