Cellevat3d® nanofiber microcarriers
Product specification overview
| Surface Area | 53 000 cm²/g |
| Material | Nanofiber-based cellulose |
| Format | Monofibers |
| Bioreactor | Stirred tank bioreactors |
| Diameter nm | 500nm |
| Animal free available | Yes |
| Typical cell density (HEK/AAV) | 2-3×106 cells/ml |
| Sterile procedure | Autoclavation |
| Scalability | From lab- to large-scale production |
| Charge | Positively charged |
| Applications | Gene therapy, PSCs, HEK based recombinant proteins, HEK based vaccines, organoid formation |
The commercial launch of Cellevat3d® nanofiber microcarriers is set for November 2024.
Cellevat3d® nanofiber microcarriers – are engineered for stirred tank bioreactors and designed, based on nanofiber cellulose monofibers to support applications such as gene therapy, pluripotent stem cells (PSCs), organoid formation, HEK-based recombinant proteins and HEK-based vaccine production.
Cellevate accelerates its launch of Cellevat3d® nanofiber microcarriers to November 2024 thanks to comprehensive validation and optimization program
Cellevate is accelerating the launch of its Cellevat3d® nanofiber microcarriers with a comprehensive evaluation, optimization and validation program, involving several partners from Europe and USA, and targeting a November 2024 release. This program focuses on validating the enhancements in productivity and process efficiency that Cellevat3d® nanofiber microcarriers offer. The aim is to optimize protocols across various scales, from laboratory to pilot and large-scale bioreactor production. The validation spans multiple reference processes with the main focus on AAV (adeno-associated virus) production.
In addition, Cellevate has already implemented the key components of a quality management system for the launch of Cellevat3d® nanofiber microcarriers starter pack in November 2024, and continues to add the remaining processes in production according to GMP and for ISO 9001 certification in the second half of 2025, when the bulk Cellevat3d® nanofiber microcarriers product portfolio will be launched.
Cellevat3d® nanofiber microcarriers designed for improved upstream productivity of viral vector biomanufacturing
Jessica Giacomoni 1, Anki Knutsson 1, Henrik Bratt 1, Jean-Christophe Bourdon 2, John Counsell 3, Luke Tappouni 3, Mikael Sommarin 4, and Linda Dexlin Mellby 1
1. Cellevate AB, Lund, Sweden, 2. Cellular Medicine, University of Dundee, Dundee, Scotland, 3. UCL Division of Surgery and Interventional Science, London’s Global University, London, UK, 4. Redoxis AB, Lund, Sweden.
Background
Gene therapy and viral vector production are associated with high manufacturing costs that restrict patient access. There is a growing demand for innovative cell culture techniques that improve productivity and scalability. Cellevat3d® nanofiber microcarriers provide a scalable solution for adeno-associated virus (AAV) production in adherent 3D culture systems.
Results
Data showed that HEK293T cells adhering to Cellevat3d® nanofiber microcarriers form homogeneous spheroids in 3D cultures with >95% cell viability and a 13-fold increase in cell density in 72 hours (Fig 1A, 1B). Scale up from ml to L scale demonstrated consistent cell growth curves, achieving yields of up to 3×106 cells/ml (Fig 1C). Further, the generated spheroids increased in size reaching an average diameter of 179 µm after four days of culture (Figure 1D).
Plasmid transfection showed high transfection efficiency of 88-94% for AAV2 and AAV9 (Fig 2A, 2B), with AAV2 yielding 1014 vg/L which was considerably higher than for 2D cultures (Fig 3A, 3B). Benchmarking performed at UCL, London, UK demonstrated a 2.7-fold increase in AAV9 vector yield compared to standard microcarriers (3C).
Cellevat3d® nanofiber microcarriers for 3D cell culturing and scale-up
Figure 1. HEK293T cell growth on Cellevat3d® nanofiber microcarriers in different culture formats and volumes. A Bright field microscope pictures of HEK293T cells adhering to nanofiber microcarriers at inoculum and growing as homogeneous 3D spheroids shown after 72h in culture. Scale bars 25 µm (left top), 50 µm (left bottom) and 100 µm (right). B Cell growth curves displaying a similar growth trend in two different cell culture formats reaching a viable yield up to 2×106 cells/ml in 3 days, while retaining viability above 95% throughout the culture. C Scalability from ml to L scale in spinner flasks with retained cell growth curves reaching a yield up to 3×106 cells/ml in 4 days. D Nanofiber microcarrier cultures generate 3D spheroids that gradually increase in size over time reaching an average diameter of 179±8 µm in 4 days (n=50 measurements per timepoint). Data are presented as either mean or mean ± SEM.
Viral vector production using Cellevat3d® nanofiber microcarriers
Figure 2. Transient transfection of HEK293T cells on Cellevat3d® nanofiber microcarriers.
A Plasmid transfection efficiency (%) measured by flow cytometry for AAV2 and AAV9 (n=3-6
biological replicates). B Fluorescence microscope images of ZsGreen-expressing transfected
3D spheroids at 72h post-transfection. Data are presented as mean ± SEM.
Figure 3. Volumetric productivity of AAV vectors using Cellevat3d® nanofiber microcarriers. A AAV2 titer of 1.5×1011 vg/ml measured by qPCR (n=4 biological replicates) from 20ml culture.
B Increased product yield (1.5-fold) over parallel AAV2 production using 2D (culture flasks) with same number of cells. C Benchmarking data showed 2.7-fold increased in AAV9 yield when using Cellevat3d® nanofiber microcarriers compared to current standard microcarriers (n=5-6 biological replicates). Data are presented as mean ± SEM.
Conclusions
Cellevat3d® nanofiber microcarriers offer a sustainable and scalable solution for upstream bioprocessing, enabling high-density 3D cell cultures with high product yields. The large surface area of the nanofiber microcarriers and the observed spheroid structures was associated with high transfection efficacy and AAV vector productivity, with a 2.7-fold increase in AAV9 yield compared to standard microcarriers.
These findings underscore Cellevat3d® nanofiber microcarriers’ potential to enhance gene therapy applications by improving yield and reducing costs.
For more detailed information, download the poster below.
Poster presented at ISCT Europe 2024 Regional Meeting, Gothenburg, Sweden, September 4-6, 2024
Pre-order
Pre-order Cellevat3d® nanofiber microcarriers starter pack today and be the first to receive it or book a meeting to learn more.
