1. การประชุมวิชาการ เรื่อง Plant Stem Cell in Cosmetics August 25, 2010
Current Research and Product Development
Using Plant Stem Cells
Malyn Chulasiri BSPharm, PhD
Director, Research & Development, SJI
Professor Emerita, Faculty of Pharmacy, Mahidol University

2. I Can’t Believe The Hype – Beauty Products With Plant Stem Cells by Aaron Saenz on June 30th, 2010
- Claims surrounding stem cell treatments are starting to get downright ridiculous - Various cosmetics lines have begun to market their goods as containing the benefit from plant stem cells - Many claims that these plant based stem cells will stimulate human stem cell growth and promote healthier younger looking skin - Plant stem cells as age-defying makeup creams? Seriously? Have we reached the point where we’re so enraptured with the possibility of human stem cell technologies that we’ll buy anything with ’stem cells’ in the product name?

3. Stem Cells
- Are indispensable for growth and development in both animals and plants
Have two properties Self-renewal : the ability to go through numerous cycles of cell division in an undifferentiated state Differentiation : the capacity to differentiate into other cell types
- May be able to repair or replace damaged tissue

4. Animal Stem Cells
♦ Have two basic types ♦ Embryonic stem cells (pluripotent) are derived from the inner cell mass of an early stage embryo are abundant in embryo and will give rise to numerous organs, tissues and cells in the body ♦ Adult or somatic stem cells (multipotent) are undifferentiated among differentiated cells in a tissue or organ generate all the cell types of the organ from which they originate – potentially regenerating the entire organ from a few cells
Injuries, illnesses and aging are inevitable parts of our lives. The human body can deal with basic injuries and illnesses in daily life by cell regeneration. However, aging and certain injuries have no cure since the tissues cannot be fully restored. This is related to the function of stem cells, the unique cells that are responsible for refreshing tissues and compensating for cellular losses

5. Animal Stem Cells
♦ Are responsible for refreshing tissues and compensating for cellular losses and are susceptible to degradation that comes with age ♦ Are rare and constantly lost in the tissues to make the tissues be less able to grow, maintain and repair
♦ Growth may be uncontrollable and cancerous cells may reproduce causing tumors
Injuries, illnesses and aging are inevitable parts of our lives. The human body can deal with basic injuries and illnesses in daily life by cell regeneration. However, aging and certain injuries have no cure since the tissues cannot be fully restored. This is related to the function of stem cells, the unique cells that are responsible for refreshing tissues and compensating for cellular losses

7. Skin Stem Cells Normal Without C/EBP-beta Differntiated cells
In normal skin (left), the stem cells at the base, shown in green, differentiate into skin cells, shown in red. In mice whose skin has
neither C/EBP-beta nor C/EBP-beta (middle), this differentiation is blocked: green-labeled stem cells appear in upper layers of skin,
and there are no differentiated skin cells (no red staining). This also happens at the initial stages of basal cell carcinomas. In skin
where C/EBP-beta is present but has lost its capacity to interact with E2F, a molecule that regulates the cell cycle (right), skin cells
start differentiating abnormally, before they have properly exited the stem cell “program” (yellow/orange). This is similar to what is
observed in the initial stages of squamous cell carcinomas, a more aggressive and invasive skin tumor
Stem cells
Stem cells

8. Stem Cells in Skin
 Epithelial skin stem cells in basal layer of epidermis and hair bulge stem cells in hair folloicle are basically found
 Epithelial skin cells are only capable of a certain number of cell divisions and then die off – they are shed constantly and require continuous duplication of new cells  Partly become fibroblasts responsible for collagen, elastin and hyaluronic acid production
A loss is detrimental to the skin (one ages skin stem cell division diminishes dramatically causing in wrinkles and thinned skin)

10. Plant Stem Cells ● Have the ability to regenerate the whole plant
(any kind of cell or tissue) ● Have a much longer life
● Have a built-in mechanism that, which causes the cell to ‘self destruct’  if damage to the DNA is detected and defective DNA will not be passed on
● Cannot make damaged or cancerous cells reproduce themselves

11. Plant Stem Cells : Formation of Callus Tissue
● A callus tissue is formed when a plant is wounded ● The normal differentiated cells in the callus tissue go back into a dedifferentiated form and become stem cells
● Mechanism is controlled by different plant hormones (auxins)

12. Stem cells can make identical copies of themselves as well as generate specialized cells
Day Day Day 6
Stem cells grown into filamants
Stem cells
Stem cells
Progenitor cell (stem cell)
Dead cells
Development into protonemal colony
Transient amplifying cell
Differentiated cell
Reprogramming of P. patens leaflet cells into stem cells and subsequent protonemata development

13. Why Plant Stem Cells for Human Use? Growth Enhancing Hormones : Auxins
- Have an essential role in coordination of many growth and behavioral processes in the plant life cycle
- The important ones include 4-chloro-indoleacetic acid, phenylacetic acid and indole-3-butyric acid
May induce the cell differentiation and regeneration of the vascular tissues when plant is wounded - Provide the ability to regulate protein synthesis (evidences showed the potential to promote collagen and elastin)
- Only found in plant stem cells - Are not present in the adult plants
□ Contain compounds that are beneficial for human health and beauty. They are  vitamins, minerals, oligoelements, growth enhancing hormones (e.g., auxins, gibberellins), cytokinins, nucleic acid and enzymes essential for plant growth
 many interesting plant substances (e.g., flavonoids) with new, powerful multi-functional and special properties e.g., detoxifying, UV stress protection □ Potentially cope with environmental stress
Plant stem cells contain the plant hormones auxin and gibberellin. Studies have shown that auxins and gibberellins are NOT present in the adult plants; they are only found in Plant Stem Cells. A lowering of oestrogen and progesterone levels leads to undesirable skin changes - delayed cell division, decline in collagen formation, thinning of the skin with a reduction in the natural moisture retaining ability and elasticity, resulting in wrinkles.

14. Plant Stem Cells : A New Ingredient
for Health & Beauty
 GMO-free
 Sustainable eco-friendly  Allows utilization of even the most rare plant species without harm
 Free of heavy metals, pesticides, and other environmental contaminants   No limitation of plant sources due to any restrictions
 Rare active molecules of interest can be concentrated up
 Can be developed exclusively to meet one’s needs
 Safe for both cosmetic and nutritional applications

15. Plant Stem Cell Technological Process (Example)
Taking away a small piece of the plan
Wounding of plant material to induce callus formation
Incubation on agar plates
Harvesting of developed callus
Cultivation until complete dedifferentiation to obtain stem cells
Transfer of the stem cells into a suspension and grow
Growing into plants

16. Plant Stem Cells : Utilized Parts as Ingredients

17. Plant Stem Cells Versus Human Skin
(for Beauty)
 Are compatible with human skin  Protect human skin stem cells
 Maintains the self renewal capacity of human skin stem cells  Increase longevity of human skin stem cells
 Replenish, regenerate and repair damaged DNA  Delay aging of essential cells  Stimulate duplication of human skin stem cells  Protect the skin from ozone/pollution damage and UV damage

18. Marketed Plant Stem Cells for Cosmetics
Origin
Trade name
Source
- Panax ginseng (Wild Korean ginseng)
- ActostemR WKGBASE - ActosomeR GINLIFT
ACT, Korea
- Cyperus papyrus (Papyrus)
- Crodarom Papyrus
Crodarom,
France
- Buddleja davidii (Summer lilac) - Centella asiatica (Asiatic pennywort) - Echinacea angustifolia (Echinacea) - Leontopodium alpinum (Edelweiss)
- Buddleja davidii Stems GTM - Centella asiatica Stems GTM - Echinacea Angustifolia Stems GTM - Leontopodium Alpinum Stems GTM
IRB, Italy
Rhododendron ferrugiineum (Swiss Alp rose) - Malus Domestica (Apple)
- Vitis Vinifera (Grape)
PhytoCellTectTM Alp Rose - PhytoCellTecTM Malus Domestica
PhytoCellTecTM Solar Vitis
Mibelle Biochemistry, Switzerland

19. Stem Cells for Cosmetics from Flower Plants
Rhododendron ferrugiineum
Increase skin cell vitality; boost epithelial regeneration; help skin to cope with climate changes
Buddleja davidii
Protect photodamage and oxidative stress; offer complete treatment for acne disorders
Leontopodium alpinum
Reduce eye contour and collagen loss; prevent photoaging
Echinacea angustifolia
Promote collagen; revitalize dull, tired skin

20. Stem Cells for Cosmetics from Fruit Plants
Malus Domestica
Preserve the youthful look and the vitality of skin
Vitis Vinifera
Protect skin stem cells against UV stress, fight photoaging
Stem Cells for Cosmetics from More Kinds of Plants
Centella Asiatica
Firm skin; improve skin tone and elasticity
Cyperus papyrus
Moisturize, soothe and energize the skin
Pinax ginseng
Reduce wrinkles; inhibit adipo-cyte differentiation

21. Edelweiss
Grows spontaneously on mountains range from Pyrenees and Alps to Himalaya Develop many defensive mechanisms in order to survive the extreme heights and protect from high level exposure to solar radiations
where the air is very clear, exposed to high UV light and extremes temperature.

22. Example of Marketed Edelweiss Stem Cells for Cosmetics Use :
Leontopodium Alpinum Stems G
From Edelweiss, the symbol of “Alpine flora” with the aid of High Tech Nature (HTN) biotechnologoy

23. HTN biotechnology is the unique process that allows the in vitro culture of plant stem cells in highly controlled conditions and enables to produce the active substances with several innate advantages :
- be able to produce substances that are rare and difficult to obtain from nature or synthesis
- secure the growing stem cells from pesticides, heavy metals and aflatoxins
- possibly obtain stems cells from endangered or protected plants without altering the ecological balance and biodiversity - supply plant products independent of climatic and geographical variability
Callus

24. Cosmetics with Ingredient from Edelweiss Stem Cell Ingredient
contains specific substances with high biological activities
- Leontopodic acids A and B, chlorogenic acid, 3,5-dicaffeoylguinic acid : antioxidant, anti-hyaluro-nidasic, anti-collagenasic activity
- Phytosterols, amino acids, polysaccharides : moisturizing and nutrients
Cosmetic Use
the ultra violet light absorbing chemicals this plant has developed from high altitude growth makes it a good additive to sun blocks. some chemicals in the plant prevent amplification of oxides, which are tied to the aging process.8. Used in high quality anti aging and anti wrinkle cosmetics.
> Reduction of eye contour wrinkle > Prevention of collagen loss
> Firming and restructuring treatments > Prevention of aging and photo-aging 24

25. Edelweiss Stem Cell Treatment25

26. For Health An Apple a Day Keeps the Doctor Away For Beauty An Apple a Day Keeps the Wrinkles Away
Currently, apple stem cell technology is popping up everywhere for beauty use

27. Apple Stem Cells as Anti-Aging Ingredient
- The current find is a rare Swiss apple, called the Uttwiler Spatlauber, which actually heals itself when punctured
- The rare Swiss apple has a renewal power that rejuvenates saggy skin - Apple stem cells help boost the production of human skin stem cells by protecting skin from stress and decreasing fine lines and wrinkles

28. Example of Marketed Apple Stem Cells for Cosmetics Use :
PhytoCellTec Malus Domestica
From Uttwiler Spätlauber, the “super apple” with the aid of modern plant stem cell technology
The fruit of Uttwiler Spätlauber apple trees drew the attention of researchers when it was discovered that the apples stayed fresh for months longer than other varieties, and did not fall from the tree or wither even if the trees themselves were dying. The apple variety is too tart to be eaten and not very pretty on the outside, but it was found to contain incredibly persistent stem cells, which were the key to its extended vitality.

29. Selecting / taking away a small piece of the plant
Wounding of plant material to induce callus formation
Incubation on agar plates
Harvesting of the developed callus
Cultivation until complete dedifferentiation to obtain stem cells
Transfer of the stem cells into liquid media for growing
Disruption of the stem cell wall to release secondary metabolites
Encapsulation of the stem cell contents into liposome
Fixing in xanthan gum gel

30. - Based on an extract of cultured apple stem cells - Formulated with nanotechnology that allows a higher encapsulation in a smaller molecule - Allows better penetration to deliver deep into the skin - Minimizes trans epidermal water loss - Enhances new stem cell production
- Protect against stem cells from UV - Released by skin heat (33oC) when is applied
the apple’s stem cells appeared to be protecting human stem cells. Like a youth tonic, they were nourishing and stimulating them, defending them from UV radiation (like a natural SPF booster), even delaying their aging and death.

31. Encapsulated nano- emulsion PhytoCellTec stem cell cream before touching the skin
Heat activated release of PhytoCellTec stem cell cream after touching the skin

32. Potentials of Malus domestica stem cell extract in protecting human umbilical cord stem cells from the impact of UV
Potentials of Malus domestica stem cell extract in increasing the number of human umbilical cord stem cells

33. Skin stem cell activity
Before PhytoCellTecTM treatment h after PhytoCellTecTM treatment

34. - Won the 2008 innovation prize for best active ingredient in Europe
- The reason it won is because its revolutionary
of stem cell technology and its ability to harness the regenerative potential of own facial stem cells to renew skin (make it look young again)

35. Cosmetics with Ingredient from Apple Stem Cells
Cosmetic Use
> Protect skin stem cells
> Increase activity of skin stem cells
> Decrease of senescence markers in fibroblasts
> Maintain the power of skin cell regeneration
> Decrease in wrinkles
Cosmetics with Ingredient from Apple Stem Cells

36. Apple Stem Cell Cosmetic Treatment
55 years old
Apple Stem Cell Cosmetic Treatment
1. Protects longevity of skin stem cells 2. Delays senescence of essential cells 3. Combats chronological aging 4. Preserves the youthful look and the vitality of your skin
Applications 1. Skin care products to protect skin stem cells 2. Real rejuvenation for face and body care
3. Innovative skin care formulations
Week Week Week Week 4
Potentials of Malus domestica stem cell extract on reducing of skin wrinkles

37. Korean Ginseng
 A medicinal herb with miraculous virtue in the high mountains and secluded valleys
 A first record of prescription of ginseng as a medicinal herb appeared in Shen-Neng Pharmacopeia (AD )
 During the Koryo Dynasty (AD ), ginseng was renowned as the best medicine with miraculous potency
 It was called a miraculous medicine, a cure-all and believed to be a miraculous medicine, a source of eternal youth 37

38. Example of Marketed Ginseng Stem Cells
for Cosmetics Use :
Actostem WKGBASE
From 100 years old Wild Korean Ginseng (rare to find, even in deep mountains) with tissue culture technology 38

39. Plant Tissue Culture Non-GMO : no genetic modification
Safety : animal free, no-infection
Eco-friendly : no extraction, no pollution
No metals : no heavy metal sources
Pesticide free : no pesticide needed
Fast growth : no need to wait 100 years for wild ginseng
Technology : tissue culture, nano-liposome
Low cost : low production cost 39

40. Genetic similarity by AFLP fingerprinting

41. Cosmetics with Ingredient from Ginseng Stem Cells
Cosmetic Use
 Collagen synthesis
 Cell proliferation
 Photo-protection
 Antioxidant
 Whitening
Contains high biological substances
- Saponins called “ginsenosides” : protopana-xadiol, triol and pleanolic acid
- Carbyhydrates : oligo- and polysaccharides, fiber, pectine
- Lipid soluble component : fat, phytosterol, essential oils, terpenoids, organic acids
- Nitrogenous compounds : Amino acids, peptide, protein, alkaloid, nucleic acid
- Vitamins & minerals
Cosmetics with Ingredient from Ginseng Stem Cells

42. Descriptions of Some Cosmetics with Ingredients
from Plant Stem Cells 42

43. Skin Care Products

44. Japan Korea Poland Oh My God Perfect Gel
IOPE Plant Stem Cell Skin Renewal Serum
Korea
Bielenda In Vitro Roslinne Komorki Macierzvste Anti-Age Day Cream SPF 10
Poland

45. USA France Italy éclos Anti-Aging Facial Cleanser Skin Prep
Rampal Latour en Provence Geneviève Patou Shampoo & Shower Gel Sweet Almond - Sage Bergamot
France
Crescina Stem Forte Re-Growth
Italy

46. IOPE Plant Stem Cell Skin Renewal Serum
IOPE's cutting edge technology brings an innovative anti-aging skin care line. Plant stem cells are the source of unlimited energy and the key to herbal growth and regeneration

47. Bielenda In Vitro Roslinne Komorki Macierzvste Anti-Age Day Cream SPF 10
Effectively repairs age-related skin damage, restores the skin's firmness and smoothness, reduces deep wrinkles, furrows and mimic lines, provides intensive hydration and protects the skin against excessive water loss, whilst restoring elasticity and glow and improving the skin's suppleness and density with its lifting effect. Formulated with a plant stem cells that naturally regenerate and renew, protect skin stem cells against ageing by delivering nutrients essential for the formation of new, young cells. The stem cells stimulate the production of collagen, reduce wrinkles and recover the skin's firmness and youth.

48. éclos Anti-Aging Facial Cleanser Skin Prep
Contains high potency plant stem cells for cellular rejuvenation. To extend skin cell longevity, cleanse deep down, and gently purify pores. Clinical results give a 100% reported decrease in wrinkle depth due to the product's tremendous ability to stimulate skin stem cells, encouraging aging skin to behave in a more youthful way.

49. Makeup Products

50. Lancôme Définicils Precious Cells Mascara
- Contains the apple extract blended with proteins and lipids for a deeply hydrating experience - The potential in stimulating the growth and health of lashes

51. Conclusion
Plant stem cells : ♥ are natural and free from many unwanted issues e.g., GMO, environmental contaminants ♥ contains substances that can be beneficial for human health and beauty
♥ Can be produced by modern technology and encapsulated for better properties
♥ Are compatible to human skin and support skin stem cells as well as help protect the skin from environmental stress
♥ are now available as the cosmetic ingredients in the market ♥ have been used worldwide in skin care and make up products

52. ACKNOWLEDGEMENTS Sunee Chongcharoenchit Siriluk Sreepan
Merck (Thailand) Ltd.
P.C. Intertrade Co., Ltd
Chemico Inter Corporation Co., Ltd
Croda Singapore PTE Ltd (Thailand Representative Office)