A comprehensive, closed cell processing system

 

The CTS Rotea Counterflow Centrifugation System offers a variety of features, all of which work together to provide a closed system able to integrate smoothly at multiple points in cell therapy processing workflows. The gentleness of counterflow centrifugation using the Rotea system helps to ensure high levels of cell recovery and viability. The single-use kits provide sterile tubing with flexible ports for input and output of media and reagents, and the intuitive software runs on an adjacent computer, allowing protocols to be viewed and modified while visually monitoring processes in real time.

Instrument

  • Compact
  • Multi-purpose
  • Easily integrates into workflow processes

Single-use kit

  • Sterile tubing assembly
  • Flexible input and output ports for connecting media and reagents

Software

  • Intuitive user-programmable interface
  • Preset protocols available
  • Suitable for GMP manufacturing

Explore the possibilities of Rotea counterflow centrifugation system:


Gentle counterflow centrifugation—how does it work?

Counterflow centrifugation (CFC) is an effective method for gently separating cells based on their sedimentation characteristics (size and density). The Rotea Counterflow Centrifugation System suspends cells in a fluidized bed by exerting a constant flow force against centrifugal forces. The fluid-suspended cells are gently concentrated without ever forming a pellet in the cone and washed efficiently at high recovery rates. Using elutriation, dead cells can be removed to optimize viability. By tuning centrifugal speed and flow rate to create an imbalance of forces, cell types of different sizes and densities can be separated.

Counterflow centrifugation using the Rotea system

Cell loading

With “balanced” g-force and counterflow parameters, input material comprising media and cells is introduced to the CFC chamber via the central tube. 

Elutriation

Larger/denser cells are captured in the fluidized bed, while smaller/less dense cells and debris pass through and are “eluted” through the top of the CFC chamber.

Media exchange/washing

Wash buffer is pumped through the fluidized bed, replacing the original media in the input product.

Note: The fluidized bed enables fast and efficient washing.

Cell concentration

Washed and concentrated cells are now recovered from the CFC chamber by simply reversing the pump and extracting the concentrate via the internal tube. 

The CTS Rotea counterflow centrifugation process is efficient yet extremely gentle on cells compared to other separation methods. The fluidized bed supports low-shear processing, enabling over 95% cell recovery while maintaining cell viability and achieving high throughput. Research shows that cell morphology and viability of cells are similar before and after counterflow centrifugation, confirming that the process minimally perturbs cells.¹

Video: See the Rotea Counterflow Centrifugation System in action

Watch to learn more about how the Rotea system and gentle counterflow centrifugal elutriation can help revolutionize cell therapy workflows.


Key features of the Rotea Counterflow Centrifugation System

Exceptional process versatility

The automated, multi-purpose CTS Rotea system is designed to fit within multiple stages of existing workflows and offers high throughput and exceptional cell recovery. These features allow the Rotea system to easily serve a variety of processes including cell separation, concentration, washing, and buffer exchange. This versatile system can process small-to-large input volumes (50 mL to 20 L) and output concentrations (5 to 500 x 10⁶ cells/mL). All cell processing is contained within a closed, sterile single-use kit, which is designed to be gentle on cells and allow for easy recovery of cells from the CFC chamber. 

Figure 1. Example CAR-T workflow. The CTS Rotea system can work with other instruments and reagents, giving you options to use it at different places within your existing process, and is suitable for wash and concentration between multiple cell therapy processing steps. Common uses in a CAR-T workflow include separating PBMCs from the apheresis product, separating immune cells such as T and NK cells, separating engineered cells such as CAR-T and CAR-NK, and cell wash.

Easy optimization with CellCam

The CTS Rotea system’s onboard CellCam (1-megapixel camera) allows cells to be viewed live in the fluidized bed during counterflow centrifugation (CFC). This live visualization allows for confirmation that the programmed CFC parameters are accurately separating, concentrating, or elutriating the cells. If needed, users can then easily modify CFC parameters to create optimized protocols for various applications.

Concentrated output volumes

The CTS Rotea system is designed to handle a wide range of input volumes from 50 mL to 20 L and produce output volumes as low as 5 mL at concentrations up to 500 million cells/mL. These highly concentrated, low-volume cell outputs contain very little buffer or carryover, making them excellent for autologous cell therapy.

Figure 2. Concentration of T cells using the Rotea system. A 1,000 mL starting volume of 500 million T cells was concentrated down to 5 mL using the CTS Rotea system.

Modular, fully enclosed cell processing system supports efficient workflows and space utilization

Due to the modularity of the system, rapid process steps such as cell separation, wash, and concentration can be decoupled from lengthy process steps like cell expansion. When scaling up, simply add as many CTS Rotea instruments as needed for each step in the workflow, reusing a single piece of equipment for multiple workflow operations. Although it can be used with biosafety cabinets and large, costly class A and B rooms, the modular, closed process minimizes the need for them. With the closed operation of the Rotea system, simultaneously process multiple batches in a shared grade C clean room using the single-use kits.


Scale smoothly from discovery through clinical and commercial manufacturing

Often, laboratory processes are not suitable for commercial manufacturing because they are not reproducible at large scale. Such processes can be difficult and costly to scale up or to transfer to a different commercial system.

The CTS Rotea Counterflow Centrifugation System can readily process a wide range of input and output volumes, making it useful not only for different tasks in the cell processing workflow but also for different stages in the development and manufacturing process. Use the system to develop, refine and automate processes, then later transition those processes seamlessly and confidently with Thermo Fisher Scientific’s software for enabling 21 CFR Part 11 compliance (Rotea SAE software or Cellmation software) to support GMP manufacturing requirements. This smooth transition also reduces the costs of moving to another system.2

Additionally, use CTS Rotea instruments for key processes to supplement, streamline, and help improve existing manufacturing workflows in fully integrated cell therapy processing systems.
 

² James D. How short-term gain can lead to long-term pain. Cell Gene Therapy Insights. 2017; 3: 271–284.


Regulatory documentation and support

Choosing a system that meets regulatory requirements is critical for the long-term success of therapeutic development and has significant implications for clinical translation and manufacturability. Investing in the development of a closed, scalable automated system prior to regulatory approval can reduce risk and potential time lost to manual systems during the progression from process development to commercialization. A Gibco CTS Rotea system supports efficient and effective cell therapy by reducing human error and allowing scientists to spend their time on critical thinking steps while the system provides reliable and reproducible results that do not sacrifice the quality of manual processing. 

 

CTS products are backed by testing and regulatory support

Cell therapy systems (CTS) products are designed to enable GMP cell and gene therapy manufacturing. They are backed by testing and regulatory support to support customers from discovery to clinical and commercial cell therapy development.

Designed to meet cell therapy regulatory requirements

CTS Rotea Counterflow Centrifuge and CTS Rotea Single-Use Kits are manufactured in an ISO 13485 facility adhering to all compliance guidelines.
 

Rotea graphical user interface software has multiple user privileges, including enhanced security, auditing, and e-signature functionality through the Rotea SAE software upgrade.
 

OPC-UA interface, an industry-standard machine communication language, enables connection to a MES/LIM or 21 CFR Part 11 compliant system.
 

The CTS Cellmation Software is powered by the DeltaV™ DCS from Emerson and has been developed following GAMP5 methods to help ensure compatibility with cGMP-compliant processes. 


Why choose the CTS Rotea system

“Rotea is exactly what the cell therapies sector has been waiting for: a flexible, single-use system that is aggressive on efficiency but gentle on cells.”

Rebecca Lim,

Hudson Institute of Medical Research

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