Adventitious agents within the cell bank
This section refers to the isolated cell line that will be used to produce the CCP. Sterility testing should be performed on the cell line to confirm the cell line is sterile prior to the proliferation production stages; this may be at the working or master cell banking stage.
Cell bank sterility testing should be conducted using a risk-based approach and must consider the biological source of the animal, geographical location of the cell source and consider pathogens on a case-by-case basis. The isolation procedures have the potential to introduce pathogens, and cell safety testing should cover any contamination likely to be introduced during this phase for example, co-cultures. A narrative and justified rationale should be provided for any testing undertaken to provide context within the application.
The application should include a justified rationale that the cell bank/starting cell material is free from any contamination that pose a safety risk. Justifications may include details of the source animal health status or may include analytical testing to confirm. All data or rationales should be included within the application alongside an evaluation of the results, where required.
Cell bank/starting material sterility should consider the following requirements:
- Biological source of the cell line sterility testing of the isolated cells should consider the biological source/donor material. Screening should consider pathogens including species specific pathogens, viruses and zoonotic viruses where appropriate. Alternate sources of contamination should be considered, for example, animal derived sera used within the isolation/production process. Screening should consider both endogenous and adventitious agents.
Where a source animal is confirmed healthy and suitable for the food chain during post-mortem inspections, analytical confirmatory testing may not be required in the isolated cell line. A scientific justification of this approach would be required.
- Mycoplasma: absence of mycoplasma should be confirmed within the cell line
- Prions: the application should acknowledge and detail the risk posed by prion disease. Where risk is considered low, the application should provide a science-based rationale.
Genomic integrity
Genome modification
It is understood that in certain cases cell lines are immortalised through genetic modification. FSA and FSS agree that because the cells do not satisfy the definition of organism due to their inability to survive and replicate outside of the bioreactor environment, the immortalised cells are not a Genetically Modified Organism (GMO). By extension, the biomass produced using such cells should not be assessed under the Genetically Modified Food and Feed Regulation (Regulation (EC) 1829/2003. However, where individual components used within the production process are derived from genetically modified organisms, and the resulting product contains residues of these GMOs, the application may be fall within the scope of the Genetically Modified Food and Feed Regulation (Regulation (EC) 1829/2003).
As this will apply to nearly all genetically modified cell lines, we anticipate most CCPs that have been produced through genetic modification will be assessed under the novel food regulations (Opens in a new window). Similarly, CCP applications will be assessed under the novel food regulations if no genetic modification has taken place.
As the techniques used in CCP production develop, it is possible that there may be cell lines that can survive outside of the bioreactor environment. This is why we recommend that if you have used genetic modification during your production process, you should contact us to discuss your application before submission, so we can advise on the correct regulatory regime. You can contact us either using our business support service (up until February 2027) or through contacting regulatedproducts@food.gov.uk.
Where genomic modification of the cell line has been used, a full and detailed description of the changes made to the cells during selection or the production processes (for example, differentiation or immortalisation) must be provided. Safety concerns of any downstream effects will need to be considered. Where appropriate, consider the use of tables, diagrams or flow charts within the application to aid description and provide narrative.
Epigenetic modification of the cell line does not change the sequence information in the genome, but it is an important regulator of genomic function, and it may give rise to changes that require safety evaluation.
Applicants should refer to EFSA 2012 Guidance on Risk Assessment of Food and Feed from Genetically Modified Animals and Derived Products for safety testing of genomic manipulated cell lines. Deviations to the guidance will require justification within the CCP application.
It is expected that the applicant will provide the relevant assessment on safety for the genetically or genomically modified cell lines, including detailed analysis of the genetic or genomic change, reasons for the change and the effect of the change on the final CCP product.
Genomic stability
Genomic instability could lead to unpredictable changes in cell behaviour which could affect product safety. Understanding genomic stability is essential for safety assessment. The application should include evidence that the cell line remains consistent and phenotypically stable throughout the production processes for example, at cell banking and at harvest.
Evidence of genomic stability may include, but is not limited to:
- Demonstrating chromosomal stability for example, karyotyping analysis.
- Monitoring cellular division for example, evidence of number of passages used to develop the cell culture for harvest, and monitoring of genetic mutation accumulations.
- Description and analysis of phenotypic stability.
- Verification of identity markers in the final harvest for example, molecular markers for differentiation
- Evidence of protein expression stability.
- DNA methylation and other epigenetic mechanisms regulating gene expression, genomic stability, and transposition.
Harmful changes due to (epi)genetic drift
Genetic drift, including epigenetic drift, in cell culture refers to random and non-random genetic changes that occur as the cells divide. Genetic drift has the potential to introduce new risks to the cell culture by changing the properties of the cells in ways that may affect safety. Monitoring of genetic stability may primarily be conducted through observation of phenotypic changes in the cell culture in comparison with the starting material (for example, cell morphology, molecular markers, exponential growth etc). Applicants should provide assurance demonstrating the absence of harmful products in the novel food as placed on market attributed to genetic drift or genetic modification for example, production of toxins or allergens. Evidence should be provided covering the entire production process from end to end.
Production processes
The production of CCPs relies on defined processes that support consistent cell growth. These processes form the foundation for ensuring traceability, quality and safety throughout production. All production processes, including all upstream and downstream processes, are subject to safety assessment to ensure that potential hazards are identified and managed.
Under Regulation (EU) 2015/2283 and Section 2.3 of the 2016 EFSA guidance for novel foods, applications should provide a description of all production processes with sufficient detail to inform evaluation of the safety. Where novel processes are used for food production, the description should consider the effect of the method on the safety of the novel food and whether new hazards are introduced.
Production process overview
To ensure that the production processes can be fully understood for safety assessment, there should be a transparent, comprehensive and detailed summary of the production and manufacturing processes used to create the novel food for placing on the market. This should include flow chart summaries and diagrams where appropriate. Flow diagrams provided should consider the confidential production processes and include a non-confidential summary. There should be a clear narrative detailing how the CCP was developed and include all processing steps and stages from initial sourcing of the cell line to harvest and storage of the CCP as placed on market.
Information to include in applications may include but is not limited to:
- Cell sourcing and sampling
- Cell isolation and cell line development
- Development and contamination control of the cell bank
- Proliferation and expansion
- Differentiation and maturation
- Harvesting
- Post-harvesting processing
- Storage of CCP material in final formulation
Industry best practices to produce food should be used. This ensures products are consistently manufactured and controlled to standards appropriate for their intended use.
Throughout the production summary, detailed descriptions of raw material quantities, equipment such as flask/bioreactor sizes, media components, volumes of media and other relevant details and quantities for the inputs is required for understanding the current scale of the process, allowing a more efficient risk assessment process. The assessment is based on the current scale of the production process as described within the application. Scale up of production methods may change the risk profile and therefore any future changes to the scale may require an application to the FSA/FSS for a change in conditions. Further details on regulatory authorisation, including scale up and re-application, will be featured in future FSA/FSS guidance.
Microbiological contamination considerations throughout production
The application should include analysis of microbiological considerations that are unique to the production methods for CCPs as novel foods, as well as microbiological considerations within food production. The application should provide a comprehensive description of the measures taken to prevent and control microbiological contamination at all stages. This should include steps to reduce cross contamination of cell lines within all stages of the production process.
Evidence should be provided showing that the potential introduction of pathogens has been considered at each stage of production. This along with the food safety management measures and Hazard Analysis and Critical Control Points (HACCP) controls in place should demonstrate how the risks are mitigated and managed.
Contamination control should include processes for decontamination of equipment and the appropriate cleaning and sanitation procedures. Evidence may include the standards for clean room conditions, description of filtration practises, environmental monitoring and controls for manual handling. Evidence of the cleaning validation should be included within the application.
Batch to batch variability
The production method must demonstrate a consistent and controlled production process with capability of reproducing repeatable batches.
The definition and justification of what constitutes an independent representative batch and how this relates to the definition of a batch in assimilated Regulation (EU) 2073/2005 must be included within the application and allow for batch-to-batch comparisons to be made effectively to assess consistency. Further supplemental guidance on independent batches can be found in FSA/FSS Allergenicity and Nutrition Supplemental guidance.
Process controls
The process controls used within the production process must show how hazards are managed and mitigated. They are steps in which a hazard can be prevented, eliminated or reduced to an acceptable level as defined in the HACCP plan. The HACCP plan must be constructed in line with best practices, in accordance with the Codex Alimentarius Commissions HACCP principles. The application should include explanations of the hazard, description of the process control points and justified measures taken to mitigate the risk and corrective measures identified under HACCP principles.
All hazards throughout the production process should be identified and mitigated, this should cover all the production steps up to and including the storage of the novel food as it is intended to be placed on market. Post-processing methods such as texturisation should be considered if applicable. Critical control points should be labelled on the production flow diagrams.
Applications should include clear information on how and why microbiological contamination could occur during each processing stage, including potential sources. Each control measure should be justified in how it is sufficient to manage microbiological risk and include supporting evidence.
The food safety management plan or HACCP must be applicable to the production methods described within the application and be suited to the current scale of production. Any scale up considerations should also be addressed.
Raw materials and inputs
Details should be provided in the application about all raw materials (including but not limited to source animal, cell line, scaffolds or cell culture media, etc.) and equipment used within all stages of production, from the cell isolation stages through to the post-harvesting processes. Applications should include CoAs or relevant data sheets to demonstrate specification compliance, such as purity. Water quality should be considered when providing raw material details. Consideration should be given to formulations and concentration of cell culture media components (including components within the media such as sera, supplement hormones, growth factors and antibiotics), cryopreservation media, scaffolds and additives etc. Raw materials and inputs into the manufacturing of CCPs may have allergenic risk, see FSA/FSS published Allergenicity and Nutrition guidance.
Any materials that contact the food during production must be appropriate for use with the food material under the proposed conditions and this evidenced within the application, including datasheets for the materials and where relevant, supporting data and an evaluation of the results. A list of contact materials must also be provided, for example, plastic tubing, bioreactor, flasks etc. Applicants are directed to EFSA guidance on Food Contact Materials (2017) for further information. It is the responsibility of the applicant to ensure that the materials used are approved food contact materials.
Where any scaffolding, beading or other structural devices are used, the sterility, raw materials and composition of the materials should be verified prior to inclusion in the production process. Where non-edible scaffolds are used, the application should clearly describe and provide evidence of how the scaffolding will be removed from the culture and detail the effect of any residuals. Where the scaffold material is edible and is intended to remain in the final product this should be addressed as part of the CCP as a novel food. Further supplemental guidance for raw material residue in the final product will be addressed in a future FSA/FSS supplemental guidance on growth media components and toxicology assessment.
Further processing
Following harvest, the CCP material may be modified using post-production processing steps such as washing, drying or purification. Where post-production steps are applied or will be applied, they must be described within the application to further detail how the novel food will be consumed under its intended use. It must be clear why each post-production processing step is carried out and any safety implications considered. If any materials are used, details of these must be included.
Where the CCP is intended to be used as an ingredient within food, there must be some consideration given to any further processing steps required to incorporate it into the final food (for example, blending, heating). An outline of these steps should be provided and include how the steps may affect the safety or stability of the novel food. Where further processing steps will be completed by a third party, within the scope of the authorisation, details of a representative example under those conditions should be used within the application.
Refer to FSA/FSS guidance for good practice in hygiene requirements
Storage conditions of the CCP at all stages post-harvest should also be described. This should incorporate details of storage duration, temperature and packaging. Details should include the controls taken to prevent contamination or degradation of the CCP.