3 PhytoCellTecTM Malus Domestica Innovation Price BSB Innovation Price for most innovative natural activeReceived during incosmetics 2008Patent pending in CH, EU, USA and Korea
4 1. Plant Cell CulturesNew cell culture technology makes it now possible to cultivate cells of endangered and rare speciesSelected plant material (leave, fruit) is wounded to induce the formation of a callus tissueThe callus of a plant consists of dedifferentiated, totipotent cells (plant stem cells)
5 The 3 Steps of PhytoCellTec Preparing a sterile plant preparationWounding the plant to induce callus tissueGrowing callus cells in liquid cultures
6 Plant Cellculture Technology (PhytoCellTecTM =PCT) Suspension cultures from plant stem cells are grown in bioreactorsMedium consists of sucrose, trace elements and plant hormones (cytokinins and auxins)Cultures grow with oxygen and without light (chlorophyll free)Batch is harvested after all sugar has been metabolizedThe cells are washed and homogenized to release the secondary metabolitesOil- and water-soluble ingredients are extracted by liposomal structures
8 Storage Ability of Apples and Skin Aging Apples with very good storage abilities stay fresh over monthsThis was an important factor for cultivar selectionModern apple cultivars are selected for intensive cultivation and for a sweet flavour since storage is no longer a problemTherefore cultivars with excellent storage abilities disappeared from the market
9 Uttwiler SpätlauberAn old cultivar is still alive in Switzerland but no longer cultivatedThe Spätlauber variety derives from the middle of the 18th centuryIt was famous for its tremendous storability without shrinkingToday this is a very rare and endangered apple variety with only a few trees left in the world
10 Uttwiler SpätlauberThe importance of the Uttwiler Spätlauber apple was honored by the release of a stamp.
15 What are Skin Stem Cells? Properties of skin stem cells:Self-renewal - the ability to go through numerous cycles and cell divisions while maintaining the undifferentiated stateDifferentiation - the capacity to differentiate into other cell types of the same tissuePrimary roles - replenish dying cells and regenerate damaged tissues
16 Adult Stem Cells in Skin In skin, different types of stem cells have been found:Type PlaceEpidermal skin stem cells Basal layer of the epidermisHair bulge stem cells Hair follicle
17 Epidermal Stem CellsStratum corneumGranular layerSpinous layerEpidermalstem cellTransientamplifyingcellsBasal layerBasementmembraneDermisDifferentiating progeny ofone epidermal stem cellStem cells are located in the basal layer where 2-7% of the cells are stem cells
19 Stem Cells and Aging Stem cells have a limited life expectation Environmental and intrinsic stress factors effect stem cellsIn aged skin the number and activity of stem cells is reducedSkin stem cells have to be protected and supported to delay preliminary aging
20 Epigenetic Factors Composition of Plant Stem Cells Plant stem cells contain similar epigenetic factors as human stem cellsPlant stem cell extracts provide important nutrients to human stem cells Plant stem cell nutrients improve the viability and healthiness of human stem cells
21 PhytoCellTecTM Malus Domestica Provides two Different Modes of Activity 1. Use of plant stem cells to protect skin stem cellsThe components of plant stem cells such as Malus domestica are rich in specific factors relevant to protect and maintain the function of skin stem cells epigenetic factors2. Use of longevity cell extract to delay skin agingThe apple Uttwiler Spätlauber is known to contain ingredients which insure longevity of cells metabolites
22 Novel Progenitor Cell Targeting Technology Biopsy: human epidermal cells ca. 6% of cells are stem cells (optically not different from normal basal cells)Medium: specifically designed, fully defined medium that mimics the physiological environment of the in vivo stem cell niche enrichment of so called progenitor cells (stem cells)3x
23 Activity Assay for Epidermal Stem Cells Stimulation of growth or over-expression of markers are not good parameters for the activity of stem cellsColony forming efficiency (CFE)Cell cultures are seeded at low density. The number of colonies formed is a value for the concentration and activity of progenitor/stem cellsDeterminationof CFE+ apple stem cell extractIncubation
24 Increase of CFE by PhytoCellTecTM Malus Domestica + 92%ResultNumber of active and healthy stem cells is increased compared to untreated culturesS-234
25 ResultPhytoCellTec apple stem cell extract enhanced CFE up to 92% compared to the control.Maintenance of the stem cell vitality of epidermal stem cells was improved.
26 Growth of Hair Follicle Hair follicles From human skin fragments (from micro dissection)Test substance PhytoCellTecTM Malus Domestica 0.2%Design Length of hair follicle increases until day 14→ after day 14 induction of senescence→ length of hair follicle decreasesS-157Day 0Day 4Day 7
27 Length of Hair Follicle Senescence of hair(stem cells) can be delayedS-157
28 PhytoCellTecTM Malus Domestica Effect on Gene Expression in Senescent Dermal Fibroblasts Cell line Normal Human Dermal FibroblastsDesign H2O2 induced senescence (2h)followed by a treatment with2% PhytoCellTecTM Malus Domestica or not (144 h)Parameters Gene profile (microarray)
29 PhytoCellTecTM Malus Domestica Gene Profile (Microarray) ↓Gen, that is down-regulated by senescence induction (see control)↑Gen, that is up-regulated or normalized in senescent cells by PhytoCellTec Malus DomesticaGenAfter H2O2Control+ 2% Malus Domestica stem cell extractcyclin B1: induces proliferation73 ↓130 ↑cyclin E1: cell cycle regulator78 ↓135 ↑p53: tumor suppressor gene63 ↓137 ↑insulin-like growth factor II: cell proliferation enhancer71 ↓117 ↑heme oxigenase 1: antioxidans enzyme89 ↓211 ↑S-194
30 In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica Analysis of wrinkle depth in the crow‘s feet area20 volunteers28 daysApplication twice dailyAgainst inital conditions
31 Capacity to Form 3D Epidermis Study designComparison of 3D epidermis formed by early passage (young) and by late passage (old)epidermal progenitor cells.Test substancewithout test substance (control)0.01% PhytoCellTecTM Malus DomesticaS-244
32 Day 14 Day 21 Day 28 Young cells (P5) (control) Old cells (P14) No cell growthNo cell growthOld cells (P14)+ PhytoCellTec MDS-244
33 ResultEpidermal progenitor (stem) cells of late passage (P14) loose the capacity to form a 3D epidermis. But when these cells are cultured with PhytoCellTecTM Malus Domestica, they retain the capacity to form a 3D epidermis similar to early passage progenitor cells.PhytoCellTecTM Malus Domestica clearly helps to conserve the progenitor cell characteristics. Also cells of late passage are able to proliferate and to undergo the complex process of stratification and differentiation.
34 In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica
35 In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica Day 282% PCT Malus DomesticaDay 0
36 PhytoCellTecTM Malus Domestica Applications Stem cell cosmeticsRejuvenation products for face and bodyFresh cell treatmentsAnti-Aging formulationsRejuvenation of hair folliclesDelay of hair aging
37 PhytoCellTecTM Malus Domestica Claims Delays senescence of essential cellsProtects longevity of skin stem cellsProtection of skin stem cellsRejuvenation of skinMaintains the power of regenerationDecrease in wrinklesCombats chronological agingWith plant stem cells to maintain the power of regenerationWith plant stem cells to delay senescence of essential skin cells
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