Plant cell culture as emerging technology for production of active cosmetic ingredients (2024)

Abstract

Plants have always been the main source for active cosmetic ingredients, having proven health beneficial effects on human, such as anti‐aging, antioxidant, anti‐inflammatory, UV‐protective, anti‐cancer, anti‐wrinkle, skin soothing, whitening, moisturizing, etc. Extracts from herbal, aromatic and/or medicinal plants have been widely used as effective active ingredients in cosmeceuticals or nutricosmetics, especially in products for topical application and skin‐care formulations. However, over the past decade, there has been an increasing interest to plant cell culture – derived active cosmetic ingredients. These are “new generation” of high quality natural products, produced by the modern plan biotechnology methods, which usually showed stronger activities than the plant extracts obtained by the classical methods. In this review, the advantages and the current progress in plant cell culture technology for the production of active cosmetic ingredients have been summarized, and discussed in details within a presented case study for calendula stem cell product development.

Keywords: Calendula officinalis L., Cosmeceuticals, Nutricosmetics, Plant cells, Plant stem cells, Polysaccharides, Tissues and organ cultures

Abbreviations

CMC

cambial meristematic cells

EGF

Epidermal Growth Factors

1. Introduction

Plants are the oldest source of natural compounds with medicinal and cosmetic properties explored by the mankind. Even nowadays, between 70 and 80% of the people worldwide relay on traditional herbal medicine to cover their basic healthcare needs 1. In fact, 11% of the existed essential drugs used in modern human medicine are still with plants origin 2. The percentage of plant‐derived natural products, applied as active ingredients in cosmetics, is much higher. In accordance with the continuous increase of world population, the global demand for natural plant ingredients is expected to continue its exponential growth 1. As result, we are already witnesses of mass overexploitation of natural habitats of some herbal and aromatic plants, placing them under threat or even leading to their extinction 3. This negative tendency is especially dangerous for medicinal plants, which in most cases are rare and endemic species. These species often grew under extreme climate, unique soil composition or specific latitude. Their populations usually have been characterized with low levels of genetic diversity, which makes these species extremely vulnerable to genetic erosion and decrease their chance to survive in case of environmental pollution 4. Moreover, plant derived active ingredients are usually very complex mix of bioactive molecules, which cannot be easily replaced by chemically synthesized analogs. To solve the above problems, plant biotechnology has provided the tool, which can secure more eco‐friendly and sustainable supply of valuable phytochemicals with health‐beneficial properties, to cover the growing demands of cosmetic industry. Over the last decade, the advantages of plant cells and tissues culture technologies have been widely explored in development of highly‐efficient platforms for more rapid production of pharmaceutically important molecules of plant origin or for heterologous expression of therapeutic proteins 5. Nowadays, there are many cosmetic products, including both cosmeceuticals and nutricosmetics, which have active ingredients, derived by plant cell culture technology. The aim of this paper is to review the recent progress in plant cell technology for cosmetic application and to provide short overview on commercialized plant cell culture – derived active cosmetic ingredients available on the market. The information in this review was retrieved by an exploratory electronic survey of plant tissue culture ‐ derived cosmetic ingredients and their technical specification datasheets, conducted by search in the technical websites of the major suppliers such as Prospector® search engine, SpecialChem, Cosmetic Design Europe, the academic search engines Scopus, ScienceDirect, Google Scholar, and PubMed, and the popular search engine Google. In addition, the main steps in development and characterization of plant cell culture – derived active ingredients are closely discussed in the presented case study for development of InnovaStemCell Calendula product.

2. Plant extracts as active ingredients in cosmetics

Plants are rich source of countless metabolites with potential application in cosmetic products. Since time immemorial, the mankind has used different plants and their extracts to create cosmetics with the aim to establishing a state of eternal youth 6. Nowadays, the plant extracts are becoming the most popular active ingredients of cosmetics due to the ever‐increasing demand for natural compounds which in addition to esthetic looks can provide additional health benefits. Development of such products, known as “cosmeceuticals”, reflects a most recent trend in the modern cosmetics and personal care industries. Indeed, plants are rich in endogenous bioactive metabolites with potential cosmetic and pharmaceutical applications 7, 8, 9. Most of these phytochemicals, such as polyphenols, phenolic acids, triterpenes, flavonoids, stilbenes, steroids, carotenoids, steroidal saponins, sterols, fatty acids, polysaccharides, sugars, peptides, etc., could be extracted with appropriate solvents and used as active ingredients present in cosmetic formulations 10. Because of this, huge numbers of plant sources have been explored by the cosmetics industry in search of innovative active ingredients which combine some specific pharmacological properties, such as antioxidant, antimicrobial, antiviral, anticancer, antifungal, anti‐inflammatory, anti‐allergy, etc., and also showing strong moisturizing, anti‐ageing, anti‐wrinkle and UV protective effects 11. However, the quality and phytochemical profiles of plant extracts varied in wide range, depending on climate, soil, latitude, seasonal factors, time of harvest, and the field management practice, which could be a challenge to standardize their activities 12. The search for novel natural phytochemicals has led to the gathering bioactive extracts not only by plants, but also from mushrooms, algae, and also, by utilization of by‐products of plant origins 13, 14, 15, 16, 17. Nowadays, with the growth in consumers’ interest to effective and safe natural products, a “new generation” of high quality bioactive phytochemicals, produced by the plant cell culture technology, have been introduced in the past decade, and now, their presence in the cosmeceuticals market have been steadily on the rise.

3. Plant cell culture technology: principles for production of active cosmetic ingredients

Plant cell culture technology is a technique for growing of plant cells under strictly controlled environmental conditions. Because plant cells are considered totipotent, they have the potential to express the full genetic machinery coded in the nucleus, and thus, they are able to produce the full spectrum of characteristic secondary metabolites, found in mother plants. Plant cells are amenable to good manufacturing practice procedures and can be easily propagated by using large volume bioreactors independently on climate or soil or field management practices 5, 18. Moreover, in vitro cultured plant cells are characterized with fast growth, and the ability to accumulate large amount of uniform biomass for a short period of time 19, 20. This is very important advantage especially for the production of rare bioactive compounds, as resveratrol, paclitaxel or terpenoids, which are usually found in low concentrations in plants and their isolation and purification requires the processing of large amounts plant biomass 21, 22, 23, 24, 25. Additionally, plant cell culture technology offers a reliable and powerful production platform for continuous supply of contamination‐free, phytochemically uniform biomass from herbal, aromatic, medicinal, and even from rare and threatened plant species 26. The perspective to obtain natural phytochemicals by using an environmentally sustainable biosynthetic platform made plant cell culture technique exceptionally attractive for the production of active ingredients for high added values “green” cosmetic formulations 27, 28, 29, 30. It should be noted, that active cosmetic ingredients, obtained by the plant cell culture technology are popularized among the customers under the name “plant stem cells”. Here it is important to understand that the term “stem cells” used in this phrase is not always referred to real plant stem cells. The most of the existed plant cell culture production platforms are in fact developed on the basis of the use of dedifferentiated plant cells rather than on the culturing of the true plant stem cells. Dedifferentiated plant cells are obtained by dedifferentiation of already differentiated mature plant cells from different specialized tissues, whereas the true plant stem cells should be never differentiated in their life cycle. Because raised from the dedifferentiation of differentiated cells, the dedifferentiated plant cell cultures could inherit some epigenetic modifications, characteristic for the type of the tissues they have obtained from, and thus, they could be very heterogeneous in their biosynthetic and growth properties. This fact made it possible to generate almost unlimited numbers of plant cell lines with unique phytochemical profiles and growth characteristics even from the same plant, used for their initiation. For this reason, the terms as “leaf stem cells”, “meristems stem cells”, “root stem cells” “rhizome stem cells”, “flower stem cells”, “fruit stem cells”, etc. could be often seen in INCI names of active cosmetic ingredients, but in fact all of these refers to dedifferentiated plant cell cultures. On the other hand, there are plant cell technologies existed, which are developed on the basis of the cultivation of true plant stem cells – the cambial meristematic cells (CMC). These cells are isolated by cambial layer and consist only of true meristem stem cells 31, 32. Cambial meristematic cells are characterized with fast and uniform growth, lack of epigenetic modifications and ability to produce predictable yields of secondary metabolites when treated with stimulating factors such as elicitors 33. This technology have been used by the Korean company “Unhwa Corp.” to produce several active cosmetic ingredients, by using the patent protected expression platform Ddobyul®, developed on the basis of propagation of cambial meristematic cells (Table 1). Another important fact, concerning the plant cells used for cosmetics, is that the term “plant stem cells” is often equally used for active ingredients, produced by either callus cultures, cell suspensions or hairy roots. It is important to understand, that the callus cultures are plant cells, cultured on solid medium, whereas the cell suspension cultures are single plant cells or small cell aggregates cultivated under submerged conditions in liquid medium. Both callus cultures and cell suspensions could consist of dedifferentiated or true stem cells. On the opposite, the hairy roots are organ cultures, obtained by genetic transformation of plant cells 23, 34. Some of the plant cell culture technology – derived active cosmetic ingredients, currently available on the market are reviewed in Table 1.

Table 1.

Some of the most popular plant cell culture technology – derived active cosmetic ingredients, currently available on the market

Market names of active ingredientPlant speciesType of cell culture and extractsBenefitsCompany; country
Phyto‐Biotics Perilla®Perilla frutescenscell suspension extractanti‐aging, antimicrobial, soothing effectActive Concepts LLC; USA
Phyto‐Biotics Quercus®Quercus albameristematic stem cells extractantioxidant, soothing effect, antimicrobial, anti‐agingActive Concepts LLC; USA
Phyto‐Biotics Acai®Euterpe oleraceacell suspension extractantioxidant, anti‐aging, anti‐wrinkle, soothing effect, moisturizerActive Concepts LLC; USA
AKOSKY® APIUMApium graveolenscallus culture extractskin regenerationAkott Evolution S.R.L.; Italy
VITADENIA®Gardenia taitensiscallus culture extractanti‐aging, anti‐wrinkle, regenerative, repairingBiocosmethic; France
UrbanEthic®Gossypium herbaceumstem cell extractanti‐photo‐aging, protection against atmospheric pollutants and heavy metals. protection from oxidative stressBiocosmethic; France
NatureCells HydragenesisVitis vinifera cv. Verdejoliposomal complex of cell suspension extractantioxidant, moisturizer, hydrating effectInfinitec; Spain
NatureCells AntiagingVitis vinifera cv. Mencialiposomal complex of cell suspension extractanti‐aging, antioxidant, anti‐wrinkleInfinitec; Spain
NatureCells Anti Stretch MarksCentella asiaticaliposomal complex of cell suspension extractskin‐firming, anti‐stretch mark effect, anti‐inflammatory, anti‐celluliteInfinitec; Spain
InnovaStemCell Calendula WCalendula officinaliscell suspension extractanti‐wrinkle, skin regeneration, moisturizerInnova BM; Bulgaria
InnovaStemCell Calendula EMCalendula officinalisemulsified cell suspensionanti‐wrinkle, skin regeneration, deep hydrationInnova BM; Bulgaria
InnovaStemCell Rosa damascena WRosa damascenacell suspension extractskin regeneration, anti‐aging, antioxidant, anti‐inflammatoryInnova BM; Bulgaria
InnovaStemCell Rosa rugosa WRosa rugosacell suspension extractantioxidant, antimicrobial, UV protectiveInnova BM; Bulgaria
Plant C‐Stem™ Vigna RadiataPhaseolus radiatusmeristem cell culture vacuole extractanti‐aging, anti‐wrinkle, protection/repair of environmental damageInnovacos Corp.; USA
Roseroot (Rhodiola rosea L .) Plant Stem Cell ExtractRhodiola roseacallus culture extractantioxidantIn vitro Plant‐tech AB; Sweden
Milk thistle (Silybum marianum L.) plant stem cell extractatSilybum marianumcallus culture extractantioxidantIn vitro Plant‐tech AB; Sweden
Greater plantain (Plantago major L.) plant stem cell extractPlantago majorcallus culture extractantioxidantIn vitro Plant‐tech AB; Sweden
PhytoCellTec™ Malus DomesticaMalus domesticaliposomal complex of cell suspension extractanti‐wrinkleMibelle AG Biochemistry; Switzerland
PhytoCellTec™ Solar VitisVitis vinifera cv. Gamay Teinturier Fréauxcell suspension extractanti‐photo‐aging, extending skin vitalityMibelle AG Biochemistry; Switzerland
PhytoCellTec™ Alp RoseRhododendron ferrugineumcell suspension extractprotecting longevity, increasing skin vitality, protection/repair of environmental damageMibelle AG Biochemistry; Switzerland
PhytoCellTec™ ArganArgania spinosacell suspension extractimprove activity of human dermal stem cells, anti‐wrinkleMibelle AG Biochemistry; Switzerland
PhytoCellTec™ SymphytumSymphytum officinalecell suspension extractincrease skin renewal, rejuvenates skin epidermisMibelle AG Biochemistry; Switzerland
PhytoCellTec™ nunatak®Saponaria pumilacell suspension extractincrease skin elasticity, firmness and density, protection/repair of environmental damage, anti‐agingMibelle AG Biochemistry; Switzerland
RootBioTec HOOcimum basilicumhairy root culture extractreduce hair loss, inhibit 5α reductase activity, stimulate dermal papilla cellsMibelle AG Biochemistry; Switzerland
EGF (Epidermal Growth Factor);Oryza sativarecombinant proteins expressed by rice cell culturerestore reduced activity with age, stimulate epidermal skin cells, anti‐aging, anti‐wrinkel, wound healing, stimulation on collagen synthesis in aged skinNatural Bio‐Materials (NBM) Inc.; Korea
FGFb (basic Fibroblast Growth Factor)Oryza sativarecombinant proteins expressed by rice cell cultureanti‐aging, anti‐wrinkle, anti‐hair loss, hair growthNatural Bio‐Materials (NBM) Inc.; Korea
IGF‐1 (Insulin‐like Growth Factor‐1)Oryza sativarecombinant proteins expressed by rice cell culturegrowth support for skin cells with EGF, support young and healthy skinNatural Bio‐Materials (NBM) Inc.; Korea
KGF (Keratinocyte Growth Factor, Fibroblast Growth Factor‐7)Oryza sativarecombinant proteins expressed by rice cell culturespecific growth factor for keratinocyteNatural Bio‐Materials (NBM) Inc.; Korea
VEGF165 (Vascular Endothelial Growth Factor)Oryza sativarecombinant proteins expressed by rice cell cultureskin cell growth and support, wrinkel and skin aging improvement, wound healing after laser resurfacing or injury, hair cell growth and support, hair cycle regulation and anti‐hair lossNatural Bio‐Materials (NBM) Inc.; Korea
Foreseen Shield NopalOpuntia ficus indicapowdered leaf cellsantioxidant, anti‐wrinkle, anti‐aging, UV protective, decrease melanine spotsNaolys, France
Initial E [PT+TMG]Polianthes tuberosacallus culture extractanti‐aging, anti‐wrinkle, anti‐circle,
anti‐puffines, brightening, firming, soothing, moisturizingNaolys, France
Power Extension [HSB+R]Hibiscus syriacusleaf cell extract with rutinanti‐aging, anti‐wrinkle, antioxidantNaolys, France
All Even Sweet irisIris pallidaglycerin based leaf cell extractanti‐aging, anti‐wrinkle, firmingNaolys, France
All Fiber Booster Olive treeOlea europeaglycerin based leaf cell extractanti‐aging, restructuratingNaolys, France
All Fiber Booster Chinese hibiscusHibiscus rosa sinensisglycerin based leaf cell extractanti‐aging, restructuratingNaolys, France
All Fiber Booster Green teaCamellia sinensisglycerin based leaf cell extractanti‐aging, firming, regenerating, softening, restructuratingNaolys, France
Fiber Booster Plus Sequoia and Vitis flowerSequoia sempervirens and Vitis viniferaglycerin based leaf and flower cells extractanti‐aging, antioxidantNaolys, France
Inside Heart Egyptian blue lilyNymphaea caerulea lglycerin based leaf and flower cells extractanti‐aging, antioxidantNaolys, France
Revive Commiphora and
Rose from DamasCommiphora myrrha and Rosa damascenaglycerin based leaf cells extractanti‐aging, energizingNaolys, France
StandStill Rose from DamasRosa damascenaglycerin based leaf cells extractanti‐aging, firmingNaolys, France
New ReGeneration CocoaTheobroma cacaoglycerin based leaf cells extractanti‐aging, firming, regenerating, protective, restructuratingNaolys, France
Total Generation Sequoia and Egyptian blue lilySequoia sempervirens and Nymphaea caeruleaglycerin based leaf cells extractanti‐aging, regeneratingNaolys, France
Total Generation Curry plantHelicrysum italicumglycerin based leaf cells extractanti‐aging, regeneratingNaolys, France
Inside Light Poet's narcissusNarcissus poeticuscallus culture extractbrightening, lightening and anti‐blemish, anti‐agingNaolys, France
Bright Light Madonna lilyLilium candidumpowdered leaf cellsbrightening, regeneratingNaolys, France
LightWaves Defense [JS+M]Jasminum sambacsunflower oil based leaf cells extractprotective, anti‐aging, radiance, firming, anti‐rednessNaolys, France
Global Protect Common juniperJuniper communisglycerin based leaf cells extractprotective, anti‐pollutionNaolys, France
OxyRelax California poppyEschscholzia californicaglycerin based leaf cells extractprotective, antioxidant, anti‐agingNaolys, France
OxyRelax Cherry treePrunus cerasusglycerin based leaf cells extractprotective, antioxidant, cell relaxingNaolys, France
Smooth Lightening White roseRosa albaglycerin based leaf cells extractprotective, antioxidant, radiant, regenerating, detoxifyingNaolys, France
Sun Protect Date palmBombax costatumglycerin based leaf cells extractprotective, soothing, sunscreen, anti‐agingNaolys, France
Whole Protection EdelweissLeontopodium alpinumsunflower oil based leaf cells extractprotective, soothing, repairing, anti‐agingNaolys, France
Whole Protection Red‐flowered silk cotton treeBombax costatumglycerin based leaf cells extractprotective, soothingNaolys, France
Refine GingerZingiber officinaleglycerin based leaf cells extractastringent, firming, mattifying, moisturizing, antioxidantNaolys, France
Pure Light Chinese peonyPaeonia lactiflorapowdered leaf cellsmattifying, radiance, moisturizing, soothing, relaxingNaolys, France
Unwind Sacred lotusNelumbo nuciferasunflower oil based leaf cells extractsoothing, relaxing, radianceNaolys, France
Essential Being Indian jasmineJasminum sambacglycerin based leaf cells extractregenerating, detoxifyingNaolys, France
Full Detox Ylang YlangCananga odoratasunflower oil based leaf cells extractradiance, mattifying, regenerating, antioxidant, anti‐pollutionNaolys, France
Purify Apothecary's roseRosa gallicasunflower oil based callus extractsoothing, regenerating, radianceNaolys, France
Purify Aloe veraAloe barbadensissunflower oil based callus extractsoothing, regenerating, radianceNaolys, France
First Light Snow lotusSaussurea involucratasunflower oil based callus extractradiance, brightening, regenerating, antioxidantNaolys, France
HydraSourcing [AM+PS]Argemone mexicanaglycerin based callus extractmoisturizing, protective, regenerative, anti‐agingNaolys, France
HydraGeneration PapyrusCyperus papyruspowdered leaf cellshydrating, protective, barrier functionNaolys, France
HydraGeneration Pale roseRosa centifoliaglycerin based leaf cells extractmoisturizing, regenerativeNaolys, France
HydraSoothing Indian olibanumBoswellia serratasunflower oil based leaf cells extractmoisturizing, regenerative, antioxidant, soothingNaolys, France
Soothing Light Apple treeMalus domesticapowdered leaf cellssoothing, lightening, radianceNaolys, France
Fragile Vitis flowerVitis viniferaglycerin based flower cells extractsoothing, antioxidantNaolys, France
Fragile CottonGossypium arboreumglycerin based leaf cells extractsoothing, protectiveNaolys, France
Fragile Japanese Cherry treePrunus serrulatasunflower oil based leaf cells extractsoothing, antioxidantNaolys, France
Soothing Light ApricotPrunus armeniacaglycerin based leaf cells extractsoothing, lighteningNaolys, France
OvernightEnhance [MJ+C]Mirabilis jalapaglycerin based callus extractradiance, energizing, repairing, detoxifying, antioxidantNaolys, France
Balancing Energy Asian ginsengPanax ginsengsunflower oil based leaf cells extractenergizing, antioxidantNaolys, France
Light&Energy Coffee and SaffronCrocus sativus and Coffea arabicasunflower oil based callus and leaf cells extractenergizing, antioxidant, radianceNaolys, France
Full Energy VanillaVanilla planifoliasunflower oil based leaf cells extractenergizing, antioxidant, regeneratingNaolys, France
Splint&Slim Great bougainvilleaBougainvillea spectabilispowdered leaf cellsfirming, slimming, lipolitical, anti‐agingNaolys, France
EtHAIReal PeppermintMentha piperitasunflower oil based leaf cells extractregulating, soothing, antioxidant, lighteningNaolys, France
Healthy Shine LilacSyringa vulgarisglycerin based leaf cells extractrestoring shine and damaged hair, soothing, energizing, protectiveNaolys, France
3HC Hair Stimulation ComplexVitis viniferamix of meristem plant cell culture derived active ingredients and hydrolyzed plant extractsincrease the lifespan of hair and reduce hair lossPhenbiox SRL; Italy
Soy CellGlycine maxcallus culture extractanti‐photo‐aging, antiradical, anti‐wrinklesPhenbiox SRL; Italy
G‐CellVitis viniferameristem cell culture from green unripe grapes extractantioxidant, skin protectivePhenbiox SRL; Italy
P‐CellCapsicum annuumfruit meristem cell culture extractpromote cell protein synthesis, increase skin elasticityPhenbiox SRL; Italy
Citrustem™Citrus aurantium dulciscallus culture extractanti‐aging, skin conditionerProvital Group; Spain
Lingostem™Vaccinium vitis‐idaeacallus culture extractanti‐aging, anti‐wrinkle, make up treatmentsProvital Group; Spain
RASTEMLpomoea purpureacallus culture extractskin‐renewal, anti‐inflammatory, anti‐wrinkle, antioxidantRadiant Inc.; Korea
Carrot Stem CellDaucus carotacallus culture extractantioxidant, skin‐nourishing effectRadiant Inc.; Korea
Cucumber Stem CellCucumis sativuscallus culture extractskin cooling effect, moisturizerRadiant Inc.; Korea
Ginseng Stem CellPanax ginsengcell culture extractskin regenerationRadiant Inc.; Korea
Lotus Stem Cell ExtractNelumbo nuciferacallus culture extractcontrols excessive sebum secretion, absorbs sebum, maintains clean skin conditionSandream Impact LLC; USA
Tomato Callus Stem Cell ExtractSolanum lycopersicumcallus culture extractmoisturizerSandream Impact LLC; USA
Rice Callus Stem Cell ExtractOryza sativacallus culture extractantioxidant, anti‐inflammatory, whiteningSandream Impact LLC; USA
Carrot Callus Stem Cell ExtractDaucus carotacallus culture extractmoisturizerSandream Impact LLC; USA
Rose Callus Stem Cell ExtractRosa damascenacallus culture extractprotecting longevity, delaying senescence, increasing skin vitality, boosting epidermal skin cell regenerationSandream Impact LLC; USA
Grape Callus Stem Cell ExtractVitis viniferacallus culture extractantioxidant, anti‐inflammatorySandream Impact LLC; USA
Orchid Callus Stem Cell ExtractOrchis spp.callus culture extractpromote skin growth and proliferation, moisturizer, skin rejuvenation, soothing effectSandream Impact LLC; USA
Ginseng Callus Stem Cell ExtractPanax pseudoginsengcallus culture extractrestorative effect, tonicSandream Impact LLC; USA
Green Tea Callus Stem Cell ExtractCamellia sinensiscallus culture extractantioxidant, anti‐inflammatorySandream Impact LLC; USA
Marrubium Stems GX™Marrubium vulgarecell suspension extractprotective effect, pollution defense, skin conditionerSederma (Croda Personal Care); United Kingdom
Buddleja Stems GX™Buddleja davidiicell suspension extractanti‐photo‐aging, anti‐aging, protects from UV‐induced oxidative stress, anti‐inflammatory, matrix metalloproteinase activationSederma (Croda Personal Care); United Kingdom
Centella Stems GX™Centella asiaticacell suspension extractreduces skin redness, straightening capillary structure, fights against rosaceaSederma (Croda Personal Care); United Kingdom
Dermasyr 10™Syringa vulgarisverbascoside concentrate extracted from leaf cell culturereduce skin blemishes and inflammatory hyper‐pigmentation, control skin redness and balances of seborrhea, soothing, purifying, skin conditioningSederma (Croda Personal Care); United Kingdom
Echinacea Stems GX™Echinacea angustifoliacell suspension extractstimulates collagen synthesis, prevents collagen loss, reduces capillary permeabilitySederma (Croda Personal Care); United Kingdom
Gardenia Stems GX™Gardenia jasminoidescell suspension extractreduces MMP‐1 synthesis, inhibits collagenase activity, stimulates collagen synthesisSederma (Croda Personal Care); United Kingdom
Leontopod Stems GX™Leontopodium alpinumcell suspension extractprevents collagen loss, antioxidant, anti‐wrinkleSederma (Croda Personal Care); United Kingdom
Celtosome™ Eryngium Maritimum STEryngium maritimumdried stem cellanti‐aging, improving skin firmness, skin renewalSEPPIC (Air Liquide Healthcare); France
Celtosome Crithmum Maritimum STCrithmum maritimumdried stem cellskin lightening, wound healing, lightening effect, pigmentation regulation, anti‐wrinkleSEPPIC (Air Liquide Healthcare); France
Ddobyul®Panax ginsengcambial meristematic cells suspension extractanti‐agingUnhwa Corp.; Korea
Ddobyul®Taxus cuspidatacambial meristematic cells suspension extractanti‐allergicUnhwa Corp.; Korea
Ddobyul®Ginkgo bilobacambial meristematic cells suspension extractwhiteningUnhwa Corp.; Korea
Ddobyul®Solanum lycopersicumcambial meristematic cells suspension extractantioxidantUnhwa Corp.; Korea
Vita SeneBlockCitrus limonmix of cell wall peptides and sugars from somatic embryosinducing youth and longevity markers, protect DNAVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Vita iLuxCapsicum annuummix of peptides and sugars from cell culturesfirming effect, promoting bilirubin degradation, improves skin's regenerative processVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Hibiskin VitaHibiscus syriacuscell culture extractrestore colagen, improve skin barrier function, moisturizer, improves skin's regenerative processVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Mythos VitaActinidia argutamix of stem cell culture derived extract and hydrosoluble plant fruit extractanti‐photo‐aging, protect DNAVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Vita NovaLotus japonicusmix of cell wall peptides and sugars from somatic embryosextends cellular vitality and longevity, anti‐aging, reactivate skin's natural repair processVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Vita Genesis WhiteBrassica rapamix of cell wall fraction and hydro‐ethanolic extract derived from hairy root culturesinhibits pigmentation process, reduce melanin synthesisVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Vita FreezeSolanum lycopersicummix of peptides and sugars from cell culturesprevent premature skin ageing, improve skin elasticity, extend cellular vitality and longevityVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Daphne VitaSenseDaphne odoracellcell suspension extractstrengthens skin protecting barrier, accelerate healing of cutaneous micro‐lesions, reduce skin inflammationVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
VitaLightCirsium eriophorumcell suspension extractreduce sebum production, reduce skin inflammation, increase skin desquamationVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
VitaShapeColeus forskohliicell suspension extractanti‐fat properties, anti‐oedema effect, antioxidant, detoxifying capacityVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
BerryFlux VitaRubus idaeuscell culture extractmoisturizer, hydrating effect, improve production of ceramidesVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
FicuCell VitaOpuntia spp.cell culture extractprotect extracellular matrix, extend cellular longevity, delay premature skin ageing, improve skin firmness and elasticityVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
DoliCos PhotoProtectDolichos spp.cell culture extractUV protective,detoxifie cells, soothing effectVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Cell PulseCoffea spp.cell culture extractantioxidant, cell energiser, anti‐wrinkleVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
Lycoskin DefenceSolanum lycopersicumwater‐soluble fraction from cell culturesextend cellular vitality, protect DNA, radiance effectVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
BioNymph PeptideNicotiana sylvestrismix of peptides and sugars from cell culturesantioxidant, extend cellular vitality and longevity, anti‐wrinkle, improve skin smoothnessVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
CellintegrityRubus idaeusconcentrated water‐soluble fraction from cell culturesantioxidant, anti‐inflammatory, protect DNA, enhance cellular longevity, soothing effect, strengthening actionVitalab s.r.l. (Arterra Bioscience s.r.l); Italy
SENSIA CAROTA PRCFDaucus carota sativalysate of root stem cells cultureanti‐aging, anti‐inflammatory, antioxidantVytrus Biotech; Spain
LUMINIA GRANATUM PRCFPunica granatumpowdered plant stem cellsantioxidant, reduces hyper‐pigmentation, enhances skin radiance and glowVytrus Biotech; Spain
ARABIAN COTTON PRCFGossypium herbaceumlysate of plant stem cells culturesoothing, anti‐inflammatory, antioxidant, photoprotection, regenerationVytrus Biotech; Spain
TURMERIA ZEN PRCFCurcuma longaglycerin based lysate of rhizome cells cultureanti‐stress wrinkles, emotional hydration manager, modulator of Brain‐skin connectionVytrus Biotech; Spain
SARCOSLIM RE‐SHAPE PRCFSarcocapnos crassifolia,lysate of callus cultureskin regeneration and repairVytrus Biotech; Spain
CAPILIA LONGA PPFCurcuma longaplant growth factor peptides ‐ concentrated secretome of totipotent rhizome cellshair follicle regeneration, hair growth re‐activationVytrus Biotech; Spain
CENTELLA REVERSA PPFCentella asiaticaplant signaling peptides ‐ concentrated secretome of totipotent petioles cellsrebuilding the core skin structureVytrus Biotech; Spain

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3.1. Plant stem cells

Plant cells can be propagated and used for continuous supply of fresh plant biomass for cosmetic formulations. However, it is of great importance to understand that we cannot introduce the entire plant cells in cosmetic products and keep them alive as active ingredient there 30, 35. Plant cells are extremely sensitive to environmental factors, nutrient medium composition, osmotic and mechanical (shear) stress, gas exchange and oxygen supply, temperature, light, ionic strength and water content, and thus, they can survive neither during cosmetic products preparation, nor during storage or application of cosmetics on skin. Even a delivery system, able to maintain and supply live stem cells could be developed, the size and the specific structure of plant cells will not allow them to attach or penetrate the skin surface. Because of all discussed concerns, the plant stem cells are rather used as a raw material for preparing different types of extracts, which then could be included in cosmetic formulations as active ingredients. However, there are various products, available on the market, which are based on a whole dried plant cells and standardized on the cells count per gram of active ingredient (Table 1). Such examples are the anti‐aging active ingredient Celtosome™ Eryngium Maritimum ST (based on sea holly cell culture with 100 million cells/g of active ingredient), and the tyrosinase inhibitory skin lightening active ingredient Celtosome™ Crithmum Maritimum ST (based on rock samphire cell culture with more than 1 billion cells/g of active ingredient) (Table 1).

3.2. Plant stem cells extracts

Most of the available active cosmetic ingredients, obtained by plant cell culture technology are marketed in the form of different extracts (Table 1). In contrast to the plant – derived extracts, the extracts obtained from plant cell cultures can be easily standardized, and perfectly complying with the strict safety requirements that the high end cosmetic market constantly demands 35. Plant cell extracts are free of pathogens, agrochemical, toxic substances, allergens and pollutants, because they are produced under controlled conditions, complying with the procedures of good manufacturing practice. Depending of the type of used solvent, the plant cell extracts used in cosmetics could be contingently divided to liposoluble (extracted with oils) and hydrosoluble (extracted with glycerin) extracts, dried extracts (conditioned with maltodextrin), plant cell wall extracts (rich in peptides and sugars), nanoemulsions or suspension extracts 35. However, most of the existed extracts have been developed on the basis on extraction of target compound or group of closely related bioactive compounds, and thus, not the entire health beneficial potential of the extracted plant cells could be utilized. Some exceptions could be found, where the entire plant cells were freeze dried and powdered for direct application in cosmetic formulations (see some products offered by “Vytrus Biotech” and “Naolys”, Table1), or the entire cell suspensions have been, emulsified or encapsulated in liposomal complex (see some products offered by “Innova BM” and “Mibelle AG Biochemistry”, Table1) by using high‐pressure homogenizers.

3.3. Molecular farming for production of recombinant proteins

Plants are excellent production matrixes for expression of recombinant proteins with important pharmaceutical properties 36. This powerful recombinant protein expression technique, known as “molecular farming” have been widely used for the production of vaccines 37, 38, 39, cytokines 40, and even for production of therapeutic protein for human use 5, 41, 42. The classical molecular farming technology is based on genetic modification of plants for recombinant protein expression. This could be realized by permanent integration of foreign genes into the host DNA to generate stable transgenic lines, or by transient transformation of intact plant leaves 43. However, both methods requires growing of whole plants under strictly controlled environmental conditions into contained greenhouses, complying with the strict regulatory standards 36. By using such technology, The Iceland Company “BIOEFFECT” has applied for the first time a plant‐based transgenic platform for large‐scale production of Epidermal Growth Factors (EGF) for cosmetic use, expressed in genetically engineered barley seeds. The company offers a wide range of skin‐care products containing this cellular activator, which contribute for healthier and younger‐looking skin. However, the recent interests in molecular farming have been focused on adaptation of plant cell culture technology for the production of recombinant proteins. This technique offers sustainable and continuous heterologous proteins production by plant cells growing under precisely controlled micro‐environmental in vitro conditions in bioreactors. Cultivation of genetically engineered plant cell cultures have been recognized as much powerful expression platform, when compared to the classical molecular farming techniques, relying on agricultural‐scale production by growing of genetically transformed plants 18, 41. Recently, the Korean company “Natural Bio‐Materials (NBM)”, has launched series of active cosmetic ingredients containing growth factors (Epidermal Growth Factor, Basic Fibroblast Growth Factor, Insulin‐like Growth Factor‐1, Keratinocyte Growth Factor, Fibroblast Growth Factor‐7, and Vascular Endothelial Growth Factor), expressed by recombinant rice (Oryza sativa L.) cell cultures (Table 1). However, even that the human recombinant proteins, produced by this technology have undeniable advantages and are characterized with high level of purity, animal‐free, virus‐free, bacterial‐free and exotoxin‐free production, there are still some consumers which have concerns to use such products because they are expressed by genetically modified organisms.

4. Case Study: Initiation, growth, phytochemical profile and physicochemical characteristics of exopolysaccharides in INNOVA StemCell Calendula

Calendula (Calendula officinalis L.), known as “marigold”, has been widely used in traditional herbal medicine and skin care cosmeceuticals for topical application 11. The pant was shown to be rich in phenolic acids, flavonoid, triterpenes, carotenoids, aroma compounds and unique mix of polyunsaturated fatty acids 44, 45, 46. Because of its high therapeutic value and the proved cosmetic effects, the Bulgarian company “Innova BM” has developed and released on the market two high quality active cosmetic ingredients, based on Calendula dedifferentiated cell culture (Table 1). The development of these products is schematically presented on Fig. 1. The technological steps include screening of calendula plants with superior phytochemical profiles, selection, sterilization and cultivation of plant explants on callus induction medium, selection of friable cell lines with appropriate phytochemical profiles, initiation of liquid cell suspension culture and optimization of cultivation conditions and nutrient medium composition. The optimization step is critical in our technology, since a significant increase in biosynthetic potential and accumulated biomass of selected cell line can be achieved (Table 2, Fig. 2). After optimization, the selected line was scaled‐up to large scale cultivation in stirred tank bioreactor. The produced cell suspension (cells and culture liquid) was then processed by high‐pressure homogenizer to produce glycerin extract (50 % wt.) or calendula emulsion (75 % wt. cell suspension) 47. The produced active ingredients have been found to have superior moisturizing, anti‐wrinkle, hydrating and regenerative effects, when applied on skin. These effects are due to the high content of secreted exopolysaccharides, during cultivation of calendula cells (Fig. 2D). The exopolysaccharides have been identified to belong to pectin type. The crude exopolysaccharides fraction contains 879 μg/mg neutral sugars and 50 μg/mg proteins. The unique combination of polysaccharides and peptides made this exopolysaccharide fraction almost perfect for application as emulsifier in cosmetic products for topical application. The potential synergistic interactions, when applied with other popular emulsifiers, as well as their emulsion stabilization properties are presented in Table 3. The polysaccharides have molecular weight of 6.7 × 104 Da and contain 413μg/mg uronic acids. The full monosaccharide composition of exopolysaccharide fraction was determined as: glucuronic acid (13.6μg/mg), galacturonic acid (399.7 μg/mg), glucose (185.5μg/mg), galactose (179.9μg/mg), rhamnose (178.9μg/mg), arabinose (166.7μg/mg), fucose (0.6μg/mg) and mannose (4.7 μg/mg). The presence of this exopolysaccharides fraction incorporated into Innova StemCell Calendula products, in combination with bioactive compounds from the released cell content (Table 2) made these products unique and one of the kind in the market of active cosmetic ingredients.

Figure 1.

Plant cell culture as emerging technology for production of active cosmetic ingredients (1)

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Table 2.

GC‐MS profiles of Calendula officinalis stem cell suspension before and after optimization of nutrient medium composition 47

MetabolitesCalendula cell suspension before optimizationCalendula cell suspension after optimization
μg/g dry biomassμg/g dry biomass
Amino acids
Glycine62.77106.74
Serine21.09175.87
L‐Proline73.08162.56
L‐Aspartic acid10.83138.41
L‐Glutamic acid161.68274.91
L‐Asparagine19.19146.38
L‐Glutamine88.03224.10
Organic acids
Succinic acid6.40130.88
Fumaric acid7.23172.30
Malic acid5.41149.20
GABA50.02184.38
Pyroglutamic acid20.41115.51
Saturated and Non‐saturated Fatty Acids
n‐Tetracanoic acid (Myristic acid) (C14:0)26.6883.07
n‐Hexadecanoic acid (Palmitic acid) (C16:0)183.91178.69
9‐(Z)‐Hexadecenoic acid (Oleic acid) (C 18:1)39.3395.36
Octadecanoic acid (Stearic acid) (C 18:0)197.61192.01
Sterols
Campesterol59.2759.07
Stigmasterol65.3489.14
β‐Sitosterol94.82112.77
β‐Amyrin12.1377.46
Phenolic acids
Quinic acid13.4981.92
Caffeic acid62.05
trans‐Cinnamic acid170.39
p‐Coumaric acid67.63

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Figure 2.

Plant cell culture as emerging technology for production of active cosmetic ingredients (2)

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Table 3.

Synergistic effect of exopolysaccharides, released by Calendula officinalis stem cell suspension in relation to emulsion stabilizing properties in mixtures with cellulose gum, xanthan gum, guar gum, and sodium‐alginate

Model emulsion systems Oil/Water (1:1 v/v) with:Light microscopyTurbidimetric method
Emulsifying activity index (EAI), m2/gEmulsion stability index (ESI), min
0.6 % Calendula Cell Suspension ExopolysaccharidesPlant cell culture as emerging technology for production of active cosmetic ingredients (3)175.016.8
0.6 % Calendula Cell Suspension Exopolysaccharides And 0.3 % Cellulose GumPlant cell culture as emerging technology for production of active cosmetic ingredients (4)38.413.9
0.6 % Calendula Cell Suspension Exopolysaccharides and 0.3 % Xanthan GumPlant cell culture as emerging technology for production of active cosmetic ingredients (5)47.622.1
0.6 % Calendula Cell Suspension Exopolysaccharides and 0.3 % Guar GumPlant cell culture as emerging technology for production of active cosmetic ingredients (6)322.415.0
0.6 % Calendula Cell Suspension Exopolysaccharides and 0.3 % Sodium AlginatePlant cell culture as emerging technology for production of active cosmetic ingredients (7)1307.560.7

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The emulsifying activity index (EAI) and emulsion stability index (ESI) were determined according to 48. Model emulsion systems were prepared by mixing polysaccharides, water and sunflower oil for 5 min at 10 000 rpm by homogenizer (Ultra Turrax IKA T18 Basic, Germany).

5. Concluding remarks

Following the ever‐growing demand for high quality natural products of plant origin for application as active ingredients in cosmeceutical formulations, the plant cell culture technology has developed powerful production platforms which can effectively supply the customers’ needs. We are witnesses of exponentially growing number of commercialized plant cell – derived ingredients, offered on the cosmetic industry market and the diversity of utilized plant species, used for their production has continue to increase every year. In fact, the observed interest to production of plant cell ‐ derived ingredients for cosmetic needs could be correlated with the recent advance in development and commercialization of plant cell culture technology in technologically advanced countries. Moreover, the new developed techniques of gene editing, metabolite engineering and synthetic biology could have a significant impact on improvement the yields and development of tailor made cosmetic products with desired activities. The advance in molecular farming has already leaded to commercial production of rare human activator peptides, cytokines and growth factors, which are the first step in development of cosmetic products with potential to extend skin life by using the body self‐repair mechanisms. However, till now there are many open questions, concerning regulatory standards and documentation, unification of health beneficial claims and the methodology for evaluation of pharmaceutical effects, which should be answered in order to help the consumers to make the right choice of their cosmeceutical product.

Practical application

The aim of this paper is to review the recent progress in plant cell technology for cosmetic application and to provide short overview on commercialized plant cell culture – derived active cosmetic ingredients available on the market.

The authors have declared no conflict of interest.

Acknowledgments

We would like to thanks the Innova BM, Bulgaria, for the financial support, close collaboration and the shared data.

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