1. Why Allogeneic Cells? Marc S. Penn, MD, PhD, FACC
Skirball Laboratory for
Cardiovascular Cellular Therapeutics
Director, Center for Cardiovascular Cell Therapy
Director, Bakken Heart-Brain Institute
Departments of Cardiovascular Medicine,
Biomedical Engineering and Stem Cell Biology
Senior Medical Director, Emerging Businesses

2. Disclosures Company Name Current Relationship
Juventas Therapeutics CSO, Equity, Inventor
Intellect, Inc. Equity, Inventor
Cour Pharmaceuticals CMO, Equity
Prognostix, Inc. CMO, Equity, Inventor
Cardionomic, Inc. Equity, Inventor
BioHeart, Inc., SAB Member, Licensee
Oakwood Medical Ventures Venture Partner
Cardax Pharmaceuticals SAB Member
Athersys, Inc. Sponsored Research
SDG, Inc. Sponsored Research
CCO Technologies Sponsored Research
Cell Targeting Sponsored Research
Southwest Michigan Fund Consultant
MPI Research Consultant
03/2009 2

3. In Vivo Studies Confirm Favorable Immunological Profile
Immunosuppression not required with MultiStem used allogeneically or xenogeneically (human → rodent) for benefit in acute MI or stroke models
MultiStem serial administration safe → no evidence of allo-antibody / T-cell sensitization response
MultiStem appears to modulate immune / inflammatory response regionally, not globally
No interference with systemic immune response, as evaluated in ovalbumin antigen challenge studies
MultiStem homes / accumulates in sites of injury in preclinical animal models 3

4. Umbilical Cord Stem Cells
Cell Types of Interest
Totipotent
Embryonic Stem Cells
Umbilical Cord Stem Cells
MAPCs
Pluripotent
Hematopoietic SC
CD117+, CD34+
Mesenchymal SC
CD117-, CD34-, SH-1+
Cardiac Myocytes, Neurons
Monocytes, Neutrophils,
Endothelial Cells, Hepatocytes
Skeletal Myoblasts 4

5. Direct Comparison of the Effects of Allogeneic and Syngeneic MAPC
LAD ligation in Lewis Rat
Genetically marked MAPC from
Lewis Rat (syngeneic)
SD Rat (allogeneic)

6. Vascular Effects of MAPC
vWF
SMA
Overlay
PBS
Lewis SD

7. MAPC into Lewis Rat at Time of Acute MI
75%
51%
6 weeks after Acute MI 10 million MAPC or Saline at time of MI
Source of MAPC

8. Allogeneic and Syngeneic MAPC Survival and Engraftment
1 Week
6 Weeks
Lewis -> Lewis
Lewis -> SD

9. Syngeneic and Allogeneic MAPC Survival
Weeks after AMI
MAPC Engraftment
(Cells / mm2)
Lewis-> Lewis
Lewis->SD
Recipient Strain
Cells Injected at the time of AMI

10. MultiStem® Immuno-Privileged In Vitro
Mixed Lymphocyte Reaction
MultiStem does not elicit In Vitro T-Cell Response in MLR Studies
Donor 1 Cells
(Rare alloreactive T-cells in red)
Donor 2 Cells
Mixture
Allogeneic T-cell controls
Recognition of allogeneic cells causes T-cell activation and proliferation
Proliferation measurable by increase DNA synthesis
T-cells don’t react
to MultiStem (MAPC)
Self to self 10

11. MultiStem® Immunosuppress Allogeneic T Cells
MultiStem (like MSC) Exhibits Immunosuppressive Effects On MLR (human)
Dose Dependent Suppression of Allogeneic T Cell Response in MLR (Lewis rat)
MAPC (MultiStem) Suppresses Immune Response
Dose Dependent Effect 11

12. MultiStem Non-Interference with Systemic Immune Response
Healthy buffalo rats immunized IP with ovalbumin (OVA)
Antibody study
Multiple MultiStem injections and evaluation points over time
T-cell study
Single IV MultiStem injections
OVA-Antibodies: no difference between MultiStem-treated and PBS control groups
T-cell response: no difference between systemically-treated MultiStem and PBS groups
Designs
Results 12

13. Summary Disadvantages
Certain stem cell types (MAPC and MSC) can be used for allogeneic delivery without immunosuppression
Advantages for this strategy include
Delivery of cells at the time of PCI
Cells from healthy and young donors
Appear to deliver paracrine factors as well as autologous
Disadvantages
Long-term survival not yet demonstrated

14. Need for Long-term Cell Survival

15. Optical Mapping
Voltage sensitive dye: Di-4-ANEPPS

16. Arrhythmia and Cell Therapy at the Time of AMI
2 Million SKMB
Direct Injection
2 Million MSC
IV Infusion
Saline
N=8-10 animal / group
1 month after AMI

17. Effect on Electrical Stability
1 month after MI *
100 80
Percent Inducible 60 40 * 20
Ctrl No MI
SKMB
Saline
MSC
Direct Injection
IV Infusion
*p<0.05 vs. Saline Control

18. MSC Connexin Expression in vivo
Overlay
Cx40
Cx43
Cx45

19. Acknowledgements Funding Sources American Heart Association NIH
Shalom Foundation
Skirball Foundation
State of Ohio
Wilson Foundation
Commecial Collaborations
Athersys
- MAPC studies
Bioheart Inc
SKMB:SDF-1 Preclinical and Clinical trials
Juventas Therapeutics
- SDF-1 Clinical Development

20. Acknowledgements CCF Stephen Ellis, MD Ravi Nair, MD Phil Howe, PhD
James Thomas, MD
CWRU
Stan Gerson, MD
Maritza Mayorga, PhD Jing Bian, PhD
Indu Deglurkar, MD Niladri Mal, MD
Arman Askari, MD Samuel Unzek, MD
Zoran Popovic, MD Soren Schenk, MD
Nikolai Vasilyev, MD Kai Wang, MD, PhD
Ming Zhang, MD, PhD Xiaorong Zhou, MD
Mazen Khalil, MD Dominik Wiktor, MD
Yu Peng, MD
Udit Agarwal Srividia Sundararaman
Amanda Finan Nikolai Sopko
Farhad Forudi, BS
Matthew Kiedrowski, BS
Kristal Weber, BS