P Phyto Tell Pec Calus Tomestica (New Version)

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P Phyto Tell Pec Calus Tomestica (New Version)

  1. 1. PhytoCellTecTM Malus Domestica Plant stem cells for skin stem cell protection © Mibelle Biochemistry, Switzerland 2009
  2. 2. PhytoCellTecTM Malus Domestica 3 Stories 1. PCT Technologie 2. Swiss Apple Story 3. Stem Cell Cosmetics © Mibelle Biochemistry, Switzerland 2009
  3. 3. PhytoCellTecTM Malus Domestica Innovation Price • BSB Innovation Price for most innovative natural active • Received during incosmetics 2008 • Patent pending in CH, EU, USA and Korea © Mibelle Biochemistry, Switzerland 2009
  4. 4. 1. Plant Cell Cultures • New cell culture technology makes it now possible to cultivate cells of endangered and rare species • Selected plant material (leave, fruit) is wounded to induce the formation of a callus tissue • The callus of a plant consists of dedifferentiated, totipotent cells (plant stem cells) © Mibelle Biochemistry, Switzerland 2009
  5. 5. The 3 Steps of PhytoCellTec 1. Preparing a sterile plant preparation 2. Wounding the plant to induce callus tissue 3. Growing callus cells in liquid cultures © Mibelle Biochemistry, Switzerland 2009
  6. 6. Plant Cellculture Technology (PhytoCellTecTM =PCT) • Suspension cultures from plant stem cells are grown in bioreactors • Medium 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 metabolized • The cells are washed and homogenized to release the secondary metabolites • Oil- and water-soluble ingredients are extracted by liposomal structures © Mibelle Biochemistry, Switzerland 2009
  7. 7. 2. Swiss Apple Story © Mibelle Biochemistry, Switzerland 2009
  8. 8. Storage Ability of Apples and Skin Aging • Apples with very good storage abilities stay fresh over months • This was an important factor for cultivar selection • Modern apple cultivars are selected for intensive cultivation and for a sweet flavour since storage is no longer a problem • Therefore cultivars with excellent storage abilities disappeared from the market © Mibelle Biochemistry, Switzerland 2009
  9. 9. Uttwiler Spätlauber • An old cultivar is still alive in Switzerland but no longer cultivated • The Spätlauber variety derives from the middle of the 18th century • It was famous for its tremendous storability without shrinking • Today this is a very rare and endangered apple variety with only a few trees left in the world © Mibelle Biochemistry, Switzerland 2009
  10. 10. Uttwiler Spätlauber The importance of the Uttwiler Spätlauber apple was honored by the release of a stamp. © Mibelle Biochemistry, Switzerland 2009
  11. 11. Young and Old © Mibelle Biochemistry, Switzerland 2009
  12. 12. The Secret of Storage Ability • High acid and tannin concentrations • Longevity of cells • Undiscovered factors? This apple can only be used by PCT © Mibelle Biochemistry, Switzerland 2009
  13. 13. PhytoCellTecTM Malus Domestica Composition Malus Domestica Stem Cells 9.0 % Phospholipids 0.14% Glycerin 0.4 % Phenoxyethanol 1.4 % Keltrol T 1.0 % Deionized water ad.100 % CTFA/INCI Malus Domestica Fruit Cell Culture Extract (and) Xanthan Gum (and) Glycerin (and) Lecithin (and) Phenoxyethanol (and) Aqua/ Water © Mibelle Biochemistry, Switzerland 2009
  14. 14. 3. Stem Cell Cosmetics © Mibelle Biochemistry, Switzerland 2009
  15. 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 state • Differentiation - the capacity to differentiate into other cell types of the same tissue • Primary roles - replenish dying cells and regenerate damaged tissues © Mibelle Biochemistry, Switzerland 2009
  16. 16. Adult Stem Cells in Skin In skin, different types of stem cells have been found: Type Place Epithelial skin stem cells Basal layer of the epidermis Hair bulge stem cells Hair follicle © Mibelle Biochemistry, Switzerland 2009
  17. 17. Epithelial Stem Cells Stratum corneum Granular layer Spinous layer Epidermal stem cell Transient Basal layer amplifying cells Basement Dermis Differentiating progeny of membrane one epidermal stem cell Stem cells are located in the basal layer where 2-7% of the cells are stem cells © Mibelle Biochemistry, Switzerland 2009
  18. 18. Hair Bulge Stem Cells Epidermal layer Basal layer Infundibulum Sebaceous gland Outer root sheath Inner root sheath Hair shaft Hair bulge stem cells Hair follicle matrix Dermal papilla Lineages derived from bulge stem cells Differentiation pathways supplied by bulge stem cells © Mibelle Biochemistry, Switzerland 2009
  19. 19. Stem Cells and Aging • Stem cells have a limited life expection • Environmental and intrinsic stress factors effect stem cells • In aged skin the number and activity of stem cells is reduced • Skin stem cells have to be protected and supported to delay preliminary aging © Mibelle Biochemistry, Switzerland 2009
  20. 20. Epigenetic Factors Composition of Plant Stem Cells • Plant stem cells contain similar epigenetic factors as human stem cells • Plant stem cell extracts provide important nutrients to human stem cells Plant stem cell nutrients improve the viability and healthiness of human stem cells © Mibelle Biochemistry, Switzerland 2009
  21. 21. PhytoCellTecTM Malus Domestica Provides two Different Modes of Activity 1. Use of plant stem cells to protect skin stem cells The 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 factors 2. Use of longevity cell extract to delay skin aging The apple Uttwiler Spätlauber is known to contain ingredients which insure longevity of cells metabolites © Mibelle Biochemistry, Switzerland 2009
  22. 22. Protection of Stem Cells against UV Damage – Study Design Cell type Umbilical cord blood stem cells Study design Incubation for 24h with PhytoCellTec Malus Domestica Extract, UV radiation of cell cultures Control Determination of living cells MTS assay 48h after UV radiation + 0.1% PhytoCellTecTM Extract © Mibelle Biochemistry, Switzerland 2009
  23. 23. Protection of Stem Cells against UV Damage – Results PhytoCellTec Malus Domestica improves the vitality of stem cells Increased protection S-142 against UV stress © Mibelle Biochemistry, Switzerland 2009
  24. 24. Novel Progenitor Cell Targeting Technology • Cells: human epidermal stem cells (primary human keratinocytes) from a biopsie 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 keratinocyte progenitor cells (stem cells) 3x © Mibelle Biochemistry, Switzerland 2009
  25. 25. Activity Assay for Epidermal Stem Cells Stimulation of growth or over-expression of markers are not good parameters for the activity of stem cells • Colony 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 cells Determination + apple stem cell extract of CFE Incubation © Mibelle Biochemistry, Switzerland 2009
  26. 26. Increase of CFE by PhytoCellTecTM Malus Domestica 100 + 92% Increase of the CFE compared to control in % 80 60 40 Result Number of active and 20 healthy stem cells is increased compared to S-234 untreated cultures 0 0.01% 0.04% Concentration of Malus Domestica stem cell extract © Mibelle Biochemistry, Switzerland 2009
  27. 27. Result PhytoCellTec 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. © Mibelle Biochemistry, Switzerland 2009
  28. 28. 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 decreases S-157 Day 0 Day 4 Day 7 © Mibelle Biochemistry, Switzerland 2009
  29. 29. Length of Hair Follicle Control 0.2% Malus Domestica stem cell extract 20 10 Change in length compared to day 14 in % 0 -10 -20 -30 -40 -50 -60 Senescence of hair S-157 (stem cells) can be delayed -70 Day 16 Day 18 Day 20 © Mibelle Biochemistry, Switzerland 2009
  30. 30. PhytoCellTecTM Malus Domestica Effect on Gene Expression in Senescent Dermal Fibroblasts Cell line Normal Human Dermal Fibroblasts Design • H2O2 induced senescence (2h) followed by a treatment with 2% PhytoCellTecTM Malus Domestica or not (144 h) Parameters • Gene profile (microarray) © Mibelle Biochemistry, Switzerland 2009
  31. 31. 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 Domestica Gen After H2O2 After H2O2 Control + 2% Malus Domestica stem cell extract cyclin B1: induces proliferation 73 ↓ 130 ↑ cyclin E1: cell cycle regulator 78 ↓ 135 ↑ p53: tumor suppressor gene 63 ↓ 137 ↑ insulin-like growth factor II: cell proliferation enhancer 71 ↓ 117 ↑ heme oxigenase 1: antioxidans enzyme 89 ↓ 211 ↑ S-194 © Mibelle Biochemistry, Switzerland 2009
  32. 32. In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica • Analysis of wrinkle depth in the crow‘s feet area • 20 volunteers • 28 days • Application twice daily • Against inital conditions © Mibelle Biochemistry, Switzerland 2009
  33. 33. In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica 2% PhytoCellTec Malus Domestica 16 12 Decrease of wrinkle depth in % 8 4 S-186 0 Day 14 Day 28 © Mibelle Biochemistry, Switzerland 2009
  34. 34. In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica S-186 Day 28 Day 0 2% PCT Malus Domestica © Mibelle Biochemistry, Switzerland 2009
  35. 35. PhytoCellTecTM Malus Domestica Applications • Stem cell cosmetics • Rejuvenation products for face and body • Fresh cell treatments • Anti-Aging formulations • Rejuvenation of hair follicles • Delay of hair aging © Mibelle Biochemistry, Switzerland 2009
  36. 36. PhytoCellTecTM Malus Domestica Claims • Delays senescence of essential cells • Protects longevity of skin stem cells • Protection of skin stem cells • Rejuvenation of skin • Maintains the power of regeneration • Decrease in wrinkles • Combats chronological aging • With plant stem cells to maintain the power of regeneration • With plant stem cells to delay senescence of essential skin cells © Mibelle Biochemistry, Switzerland 2009
  37. 37. PhytoCellTecTM Malus Domestica Plant stem cells for skin stem cell protection Patent pending © Mibelle Biochemistry, Switzerland 2009

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