Dr Cyndy Sanberg's presentation in Taiwan

1. Cyndy D. Sanberg, Ph.D.
President & COO

2. What are Stem Cells and
Why are They Important?
Stem cells: (U.S. National Institutes of Health 2010)
 Can develop into many different cell types during early life and
growth within the body.
 Located in many tissues, serve as an internal repair system.
 Divide without limit to replenish other cells
– When a stem cells divides, each new cell remains a stem cell or
becomes another type of cell with specialized function
(i.e. muscle cell, red blood cell, or a brain cell).

3. Types of Stem Cells
3 Types of Stem cells: (U.S. National Institutes of Health 2010)
1) Embryonic Stem Cells
2) Adult Stem Cells (Somatic)
3) Induced Pluripotent Stem Cells (iPSCs) – genetically reprogrammed
adult sc that assumes a stem-like state, 2006.

4. Importance of Adult Stem Cells
 are located in most tissues throughout the body.
 can develop into most cell types.
 maintain & repair tissue.
 support a healthy immune system.
The Problem:
As we age, our adult stem cells decrease
and lose their ability to maintain healthy
immune function.

5. Goal - Adult Stem Cell Health
1) Can we design a formulation from a
nutraceutical approach?
2) If we combine potent natural
ingredients can we create a synergistic
effect?

6. Out of 100 Natural Ingredients Tested at
Different Ratios We Discovered
Green Tea Vitamin D3
4 Ingredients with Potent Synergy:
For adult stem cell proliferation
For protection against oxidation
For memory & neurogenesis
Blueberries L-Carnosine
NT-020 Formula
U.S. Patent #7,442,394

7. NT-020 Increases
Proliferation of Human Stem Cells
Human Bone Marrow Cells Human CD34+ Cells
100 100
% over control (mean ± 1 SEM)
80 80
60 60
40 40
20 20
0 0
hGM-CSF Vit. D3
Cells were cultured in 96-well plates, and
treated with extracts for 72 hrs. Blueberry
Catechin
Assay: MTT for cell proliferation Carnosine
Green Tea
Bickford et al., 2006; Stem Cells & Dev.

8. NT-020 Increases
Proliferation of Human Stem Cells
Human CD133+ Cells
60
% over control (mean ± 1 SEM)
hGM-CSF Vit. D3
40 Blueberry
Catechin
Carnosine
20 Green Tea
0
Bickford et al., 2006; Stem Cells & Dev.

9. NT-020 Increases Neuroprotection
Primary microglial cells were grown
in culture and exposed to an
oxidative insult (H2O2).
Lactate dehydrogenase (LDH) assay
was used.
Results:
~90% of the cells in the control
groups died (10% survival).
However, in the presence of
NT-020 cell death was 40%
(60% survival).
Shytle RD, Ehrhart J, Tan J, Vila J, Cole M, Sanberg CD, Sanberg PR, Bickford PC (2007) Oxidative stress of
neural, hematopoietic, and stem cells: protection by natural compounds. Rejuvenation Res 10:173-178.

10. NT-020 Increases Neuroprotection
Primary neuronal cells were grown
in culture and exposed to an
oxidative insult (H2O2).
Lactate dehydrogenase (LDH) assay
was used.
Results:
~70-80% of the cells in the
control groups died (20-30%
survival). However, in the
presence of NT-020 cell death
was 35% (65% survival).
Shytle RD, Ehrhart J, Tan J, Vila J, Cole M, Sanberg CD, Sanberg PR, Bickford PC (2007) Oxidative stress of
neural, hematopoietic, and stem cells: protection by natural compounds. Rejuvenation Res 10:173-178.

11. NT-020
Antioxidant Properties
Animals received NT-020 or
vehicle for 2 weeks orally. Bone
marrow stem cells grown in
culture, and exposed to an
oxidative insult (H2O2).
Results:
The low dose showed a 20%
(13.5mg/kg) decrease in cell death; high
N = 5 each group
(13.5mg/kg)
(135mg/kg) dose showed a greater than
40% decrease in cell death;
compared to control.
High dose = 2 caps of NT-020
Shytle RD, Ehrhart J, Tan J, Vila J, Cole M, Sanberg CD, Sanberg PR, Bickford PC (2007) Oxidative stress of
neural, hematopoietic, and stem cells: protection by natural compounds. Rejuvenation Res 10:173-178.

12. NT-020 and Stroke
NT-020 - animals received orally for 14 days
MCAO (stroke) – on day 14
Behavioral testing – occurred prior to stroke and
the 14th day following stroke
Infarction Analysis – tissues collected the 14th day
following stroke after behavioral testing
(Yasuhara et al., 2008; Cell Transplantation)

13. NT-020 Reduces Damage Post-Stroke
Degree of Damage
Vehicle
NT-020
NT-020
Bederson Test – neurological
EBST – motor function
Vehicle
NT-020 Results: NT-020 decreases the
behavioral deficit following a stroke
and reduces brain damage.
(Yasuhara et al., 2008; Cell Transplantation)

14. NT-020 Increases Stem Cell Proliferation
Results:
NT-020 increases proliferation of stem
cells in two brain regions:
1) Subventricular Zone (SVZ) - a stem
NT-020
cell rich zone.
2) Striatum - a target area for the
migration of proliferating stem cells.
NT-020 (Yasuhara et al., 2008; Cell Transplantation)

15. Control
NT-020
Stroke lesion
Stem Cells

16. NT-020
Enhances Memory
4theloveofanimals.com
Morris Water Maze
Richard G. Morris (2008), Scholarpedia, 3(8): 6315.

17. NT-020 Enhances
Memory
Commons.wikimedia.org/wiki/File:morriswatermaze
3 test groups:
1) Young (3 months)
2) Old (20 months)
3) Old Treated (NT-020)
NT-020 (135mg/kg/day) was given
orally to the NT-020 Old group for 4
weeks. Young and Old Control
animals were given water.
Test: Morris Water Maze, spatial
learning and memory for 5 days.
Richard G. Morris (2008), Scholarpedia, 3(8): 6315.
Acosta S.A. et al., Society for Neuroscience Abstracts (2009).

18. NT-020
Enhances Memory
Commons.wikimedia.org/wiki/File:morriswatermaze
Not all of the Old animals showed
impairment. Thus only the old
Day 5 impaired groups were compared
Cognitively
(top 40% of each group).
Impaired
Results:
NT-020 improved the
memory of the Older animals.
Acosta S.A. et al., Society for Neuroscience Abstracts (2009).

19. NT-020
Effects Inflammation
NT-020 Treatment Group -
135mg/kg/day orally for 4 weeks
Controls – water
Young – 4 months
Old – 21 months
Aging increases the expression of
MHC class II receptors on microglia
reflecting a rise in inflammation. Old
= 21 months
Results:
NT-020 significantly reduces MHC
Class II expression in the DG, which
is responsible for learning and
memory.
Acosta S.A. et al., Society for Neuroscience Abstracts (2009).

20. NT-020
Increases Neurogenesis
OX6 – SGZ Dentate Gyrus
Young NT-020 Treatment Group -
Young
135mg/kg/day orally for 4
weeks
Controls - water
Old Old
Results:
A significant age-related
decline in proliferating
cells (Ki67).
Old
NT-020
NT-020 increases the
Old
NT-020 number of proliferating
cells.
Acosta S.A. et al., Society for Neuroscience Abstracts (2009).

21. Conclusions
A Specific Combination of Natural Ingredients (NT-020):
 Promotes the proliferation of adult stem cells.
Has potent antioxidant properties (neuroprotection).
 Helps to minimize neural damage in an animal model of stroke.
 Increases the migration of adult stem cells to ischemic areas.
 Enhances spatial memory.
 Reduces MHC Class II expression (reduces inflammation).
 Increases neurogenesis.

22. Acknowledgements
Shinn-Zong Lin, M.D., Ph.D.
Paula C. Bickford, Ph.D.
Cesar Borlongan, Ph.D.
Doug Shytle, Ph.D.
Paul R. Sanberg, M.D., Ph.D., D.Sc.

23. USF Research Park
3802 Spectrum Blvd., Ste. 142
Tampa, FL 33612
Phone: 813-866-7818
Fax: 813-315-7995
www.naturatherapeutics.com