HEK 293one Cell Line
Adeno-associated virus (AAV) has become a cornerstone of gene therapy thanks to its safety profile, broad tissue tropism, and long-term gene expression. At the heart of efficient AAV production lies the human embryonic kidney 293 (HEK 293) cell line. Since the early 2000s, HEK 293 cells and their derivatives have become the industry standard for small- to large-scale AAV manufacturing in both academic and commercial settings. To support our proprietary AAVone production system, we have successfully developed HEK293one, a new suspension HEK 293 cell line for AAV production.
Selected with AAVone single-plasmid AAV production system
Stable single-cell suspension growth without clump formation
Fully adapted to FreeStyle F17/Viral Production Medium, PEAK/PRIME, and Pro293 media
High AAV productivity across multiple production platforms and compatibility with various AAV serotypes
AAV9: up to 5e12 vg/ml in AAVone system
AAV1/5/8: over 2e12 vg/ml in AAVone system
AAV2/6: over 1e12 vg/ml in AAVone system
- High full AAV particles compatibility with various AAV serotypes
- AAV9: ~70% full particles
- AAV1/2/6/8: >50% full particles
- AAV5: ~35% full particles
High transfection efficiency and compatible with multiple transfection reagents, especially our PEIone
Available for research use under license, cGMP cell banks are currently in preparation
HEK 293 cells were originally derived from human embryonic kidney cells transfected with adenovirus type 5 DNA. HEK293 cells are the most widely used host cell line for AAV manufacturing, spanning basic research, preclinical studies, and clinical-grade production. Their dominance is driven by a combination of biological compatibility, manufacturing efficiency, and regulatory familiarity.
1. Built-In Adenoviral Helper Functions.
HEK293 cells constitutively express adenoviral E1A/E1B genes, which supply critical helper functions required for efficient AAV replication and packaging. This eliminates the need for infectious helper viruses and enables plasmid-based AAV production systems.
2. High Transfection Efficiency
HEK293 cells are exceptionally amenable to chemical transfection (e.g., PEI-based reagents), supporting robust uptake and expression of large plasmids such as pAAVone. This property is essential for high-yield AAV manufacturing, especially for multi-gene or large helper constructs.
3. High AAV Productivity
When combined with optimized production systems (such as AAVone single-plasmid ), HEK293 cells consistently deliver:
- High vector genome (VG) titers
- Improved full-to-empty capsid ratios
- Reproducible batch-to-batch performance
This makes them suitable for both serotype screening and scaled manufacturing.
4. Adaptability to Suspension Culture
HEK293 derivatives (e.g., suspension-adapted lines) grow efficiently in serum-free, chemically defined media, enabling:
- Scalable production in shake flasks and bioreactors
- Reduced contamination risk
- Lower manufacturing cost and improved process control
- Importantly, well-optimized HEK293 suspension lines exhibit minimal clump aggregation, improving transfection uniformity and downstream processing.
5. Regulatory Acceptance and Track Record
HEK293 cells have a long history of use in FDA- and EMA-approved gene therapies, providing:
- Established safety profiles
- Well-understood impurity risks
- Streamlined regulatory review compared with novel host cell lines
Other HEK 293 Variants in AAV Production
| Cell Line | Developer/Owner | Key Features | Common Applications |
| HEK 293 | Frank Graham, 1973 (Leiden U.) | Parental line expressing adenoviral E1A/E1B genes | General gene expression studies, adenoviral vector production |
| HEK 293A | Invitrogen (Thermo Fisher Scientific) | A subclone of the HEK293 cells with a relatively flat morphology | High-efficiency transfection, general AAV production |
| HEK 293F | Invitrogen (Thermo Fisher Scientific) | Adapted for serum-free suspension culture | Large-scale AAV/protein production in bioreactors |
| HEK 293FT | Invitrogen (Thermo Fisher Scientific) | Fast-growing 293T clone with robust transfection efficiency | Rapid transient transfection, scalable AAV production |
| HEK 293H | Invitrogen (Thermo Fisher Scientific) | Optimized for high-density adherent growth | Improved viral yields under adherent conditions |
| HEK 293S | ATCC/NIH | Original HEK293 line adapted to suspension in DMEM | Scalable protein production, glycosylation studies |
| HEK 293T | DuBridge et al., 1987 | Expresses SV40 large T antigen | High-efficiency AAV, lentivirus, retrovirus production |
| Expi293F™ | Thermo Fisher Scientific | Commercial, high-density suspension HEK 293 derivative | Ultra-high yield recombinant protein |
| VCP2.0 | Thermo Fisher Scientific | A clonal cell line derived from the HEK293F | High-yield AAV production without traditional pHelper plasmid |