The InnCoCells project is developing innovative production processes for cosmetic ingredients using plant cell cultures, aeroponic systems, and whole plants.
Plants produce a remarkably diverse array of metabolites with bioactive properties. Many of these molecules are used as drugs, others as dyes or chemical building blocks, and some can be used as cosmetic ingredients. Looking at trends in the cosmetic industry, we can see that consumers are primarily interested in efficacy – they want cosmetic ingredients with real and scientifically proven effects.
They are also interested in sustainability – in other words, they want cosmetic ingredients to be produced without harming the environment. For this reason, the cosmetic industry is looking for innovations in the area of environmentally beneficial and naturally sourced ingredients with evidence-based functionality.
One of the problems with sourcing cosmetic ingredients from plants is that high demand usually leads to the overexploitation of nature. This is especially true for exotic and rare species that grow in natural habitats and have yet to be domesticated.
InnCoCells, a Horizon 2020 project with 16 partners representing 11 countries across Europe, addresses this challenge by harnessing a suite of technologies for sustainable exploitation. These include the cultivation of plant cells and plant roots in bioreactors, the use of aeroponic cultivation systems, the establishment of domesticated field plots for underexploited plants, and harnessing the biochemical diversity found in plant waste and industrial side streams – an approach known as a cascade biorefinery.
These sources of bioactive ingredients are combined with innovative extraction methods and comprehensive testing for activity and efficacy to produce a panel of promising ingredients suitable for commercialisation.
Sustainable technology platforms
Plants comprise millions of cells, each carrying the genetic information needed to make whatever molecules the whole plant can make. If a particular plant species produces a useful cosmetic ingredient, then it should be possible to grow the cells in a bioreactor and persuade them to make that ingredient too. This is not always straightforward because plants contain differentiated cells, meaning they are specialised to perform certain functions.
In contrast, cells grown in a bioreactor tend to stay in a basic undifferentiated state. It is possible to induce undifferentiated cells to make cosmetic ingredients by altering the culture conditions.
For example, plant cells behave differently when grown in the dark or exposed to different types of light, or the cell culture medium can be supplemented with components that activate particular metabolic pathways.
The InnCoCells project is testing cell suspension cultures derived from many different plants to see if we can produce the molecules that make the best cosmetic ingredients. This is achieved by cutting off some plant tissue (known as an explant) and placing it in a medium that causes the cells surrounding the cut to become undifferentiated. The resulting tissue is known as a callus, which can give rise to single cells growing in suspension or differentiated plant tissues (even whole plants) depending on the hormones and other components in the medium.
An alternative to single plant cells in suspension is whole organ cultures, such as hairy roots. A natural bacterial pathogen that lives in the soil colonises plant roots, causing the roots to grow extensively and produce many hairs. The bacteria use this as a means to survive, but because roots tend to produce various metabolites, the hairy root system can also be exploited to make cosmetic ingredients.
Both cell suspension cultures and hairy roots are sustainable because they can be produced from a single explant and will grow continuously, so no plants need to be harvested from their natural habitats.
The other systems used in the InnCoCells project involve whole plants. Aeroponic systems grow plants in racks, with the roots dangling beneath rather than being in soil. The exposed roots can be harvested to extract cosmetic ingredients, or the droplets extruded from roots can be recovered while leaving the roots intact, a process known as plant milking.
We are also growing underutilised plants in the greenhouse and field, allowing them to be harvested like crops rather than destroying plant populations in natural habitats. This domestication process can sometimes alter the plants and reduce the yields of cosmetic ingredients, so we are carefully testing the growth conditions to ensure that target metabolites are still produced in useful quantities.
All these different plant-based platforms are described as upstream production technologies because they generate biomass that contains the products of interest. Various industry side streams and waste (e.g. from the olive industry) are also being screened for valuable ingredients.
The project also has a suite of downstream processing technologies to extract ingredients and keep them stable. One of the key innovations of the project was to start off with some plant-based systems already working on a large scale and therefore ready for immediate extraction.
In contrast, others were only tested at the laboratory scale, and still others were not tested at all. This provided a range of upstream platforms at different development stages at the beginning of the project, allowing us to test extraction methods immediately and also begin functional tests on the first set of ingredient candidates, using a wide range of methods to identify cosmetic activities such as anti-inflammatory, antimicrobial and healthy ageing effects.
The InnCoCells project has already achieved its initial objectives
As this article goes to press, the project has reached the halfway stage and has achieved all its initial objectives. We began by selecting more than 50 plant species to work on, spanning the three different scales and development stages mentioned above.
One important constraint was that all the plants must be in the CosIng and IECIC 2021 databases (which list species approved for cosmetic use) and adhere to the Nagoya protocol for access and benefit sharing. As the project has progressed, we have added a number of additional promising species as well as other resources such as industry side streams.
Accordingly, more than 60 new lines have been tested – meaning plant cell suspension cultures, hairy roots, and whole plants grown in aeroponic systems or the greenhouse/field. Lines with the most promising growth properties have been assessed for their ability to produce bioactive cosmetic ingredients under diverse conditions. In nature, plants tend to prioritise either growth or the production of metabolites. In the laboratory, it is possible to achieve both processes simultaneously using physical, metabolic, and genetic tools.
Many of the cell lines that were initially only grown in flasks have been scaled up, some to 40L and a few to 500L. Similarly, some hairy root cultures that started off at the laboratory scale are now scaled up to 30L. Maintaining the optimal production of cosmetic ingredients at these larger scales is challenging, but the InnCoCells team has worked hard to achieve this. We have also scaled up several species in aeroponic systems, starting on a surface of 2m2 but now increasing to 100m2.
Large-scale plots of chrysanthemum, geranium and lilac have also been established in the field. We have tested the extraction of active compounds from many of these plants and cell/tissue cultures on a small scale to establish the best processing parameters that maintain product stability, including physical and enzymatic pre-treatment steps.
So far, the ingredients produced by 15 cell lines and several industry side streams (ginger, chicory and olive waste) have been analysed in detail by mass spectrometry to detect and identify potentially bioactive molecules. The extracts have also been tested in comprehensive bioassays, revealing several promising ingredients that inhibit microbial growth and promote collagen synthesis.
Commercial development and communication
As well as the technical work that focuses on developing upstream production platforms, downstream processing, and ingredient testing, we have also prepared documentation relating to life cycle analysis, which provides direct and quantifiable evidence of sustainability.
We have begun the techno-economic evaluation of our lead ingredients to ensure that they meet market demands and can be produced in an economically viable manner. This required the collection of technical and regulatory data and the development of surveys to determine trends in consumer expectations.
Our pathway to commercialisation is guided by our stakeholder group of 16 organisations spanning the cosmetic industry, related research fields and consumer groups. We have also developed a rich set of resources for the dissemination of project results and communication with the public, including published research articles, presentations at conferences and exhibitions, a project website and social media presence, informative brochures, promotional videos, a podcast series, and an InnCoCells Academy featuring webinars on relevant topics that are open to the public.
The project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 101000373
Please note, this article will also appear in the sixteenth edition of our quarterly publication.
