InnCoCells has now identified promising ingredient candidates!
We have prepared hundreds of extracts from plants, cells and root cultures that have been optimized and scaled up, and we have tested them systematically for ingredients that are suitable for the cosmetics industry. The first ingredient candidates for commercial evaluation are now coming through the pipeline!
Image credit: lychees by B. Navez (CC BY-SA 4.0)
Progress after 36 months - upstream production
After 3 years, the EU-funded InnCoCells project is beginning to test the safety and efficacy of its first ingredient candidates as part of pre-commercial development, but important work is still going on in the technical work packages dealing with upstream production. We have tailored cultivation processes that optimize biomass production and the accumulation of specific metabolites, making them suitable for the most advanced plants in the InnCoCells portfolio of plant species. Elicitor compounds and genetic engineering tools that were developed for model species have also been adapted to promote growth or the accumulation of target compounds in these species. We have developed optimal growth conditions for a further 17 cell cultures and three hairy root lines, some of which have been scaled up to multi-litre bioreactors growing in the dark or with illumination. We also improved the growth and metabolite production of three aeroponic systems using combinations of nutrition and elicitation. Four cell lines have been scaled up to 300 L and one to 1000 L. Technical runs have been completed for one hairy root line at the 300-L pilot scale. Two species were scaled up to 100 m2 in aeroponic units. The biomass from two fields of plants has been recovered for processing.
The picture below shows the end result when apple cells grown in a 300-L reactor are processed to produce dried biomass. The material is rich in bioactive cosmetic ingredients that can be extracted with a range of solvents. Image credit: LIST
The picture below shows other sources of biomass in the project: the roots of plants grown in aeroponic systems, in this case Pogostemon cablin (top), hairy root cultures, in this case basil (top, inset), and the floral components of plants grown in the field, in this case chrysanthemum (bottom). Image credits: PAT, EVO and ECOM
Cryopreservation and storage methods that preserve biomass accumulation and the production of metabolites were evaluated for several of the species in the InnCoCells portfolio. Reserve stocks of plant cells can be kept in the freezer and used to start new production campaigns, but only if they remain viable during long-term cold storage. This process, known as cryopreservation, must be rigorously tested. The picture below shows cryopreservation tests for cells of the tropical shrub Waltheria indica, revealing the effects of different concentrations of the cryoprotectant and the duration of the procedure. Image credit: VTT
Progress after 36 months - downstream processing, extract analysis and product testing
Our latest data show that the pre-treatment of plant material with enzymes and pulsed electric fields has a positive impact on the yield and functionality of the resulting extracts. Laboratory-scale extraction protocols have been scaled up and the stability of the extracts has been studied and documented. Further work has also been carried out to optimize the use of by-products and waste fractions, such as the aerial plant parts left over from the aeroponic cultivation system.
More than 100 hydrophilic and lipophilic extracts have been tested for a wide range of properties including anti-inflammatory, anti-aging, antioxidant and antimicrobial activities. The chemical composition of extracts showing the most promising cosmetic-related bioactivities has been assessed in detail using a combination of targeted and untargeted metabolomics. The picture below shows that even small changes in culture conditions can have a big effect on the metabolites produced by plant cell cultures. The distribution of polar metabolites in chili cell cultures grown at different temperatures is shown with or without pH adjustment in a volcano plot. The red dots are metabolites that are enriched and the green ones are depleted. The InnCoCells project aims to find culture conditions that favour the production of bioactive metabolites that are suitable as cosmetic ingredients. Image credit: ENEA
Dissemination, communication and exploitation
As more ingredients come through the pipeline, we are increasingly focusing on the later stages of development, including regulatory compliance, life cycle assessment and the evaluation of product and process sustainability. This requires extensive documentation and the assessment of techno-economic viability. Regulatory compliance has been evaluated for selected ingredients, and we are working on the preparation of safety and technical data sheets, scientific dossiers, and the pre-commercial evaluation of efficacy claims. We have completed the techno-economic evaluation and life cycle assessment of an InnCoCells cosmetic ingredient derived from pilot-scale plant cell suspension cultures, including a preliminary environmental impact assessment that considers factors such as global warming potential, eutrophication, water consumption, and land use. We have now published 10 scientific articles and have presented talks and posters at more than 50 conferences. We have also raised awareness of the project at several high-profile cosmetic industry exhibitions. We have presented seven InnCoCells Academy public webinars and two more for the consortium and invited stakeholders, as well as a training workshop on plant cell cultures. Our Stakeholder Group, which provides feedback and advice, has attended multiple project meetings and has helped with the selection of promising cosmetic ingredients as well as amplifying our dissemination and communication activities. We have continued to raise awareness about the project by attending media events and by publishing brochures, podcasts, videos, updates on our website, and posts on our four social media channels.