Overview: In this webinar, Max Myers presented his work on the use of autologous bone marrow-derived stem cells injected into the cortex of rats, following a stable stroke. Max also demonstrated its lab’s findings and talked about the Aspect Imaging M7 compact MRI system as it relates to its use in this project. Key Points: The critical use of stem cells in stroke research Overcoming the blood-brain barrier via intracerebral injection of stem cells The introduction of stem cells led to improved functional recovery following an ischemic stroke How MRI can contribute to the understanding of treatments following stroke

1. Intracerebral Transplantation of Autologous
Bone Marrow Stem Cells Produces Functional
Recovery in Rats with Long-Term Stable Strokes
Max Myers
Research Technician, Iacovitti Laboratory,
Thomas Jefferson University

2. Stroke is the 5th Leading Cause of Death in the United States
● Every 40 seconds, someone in the United States has a
stroke. Every 3.5 minutes, someone dies of stroke
● Every year, more than 795,000 people in the United
States have a stroke. About 610,000 of these are first
or new strokes
● About 185,000 strokes—nearly 1 in 4—are in people
who have had a previous stroke
● About 87% of all strokes are ischemic strokes
● Stroke-related costs in the United States came to
nearly $53 billion between 2017 and 2018

3. How stroke is currently treated
● Throughout the past two decades, the management of ischemic stroke has progressively changed, shifting from
an approach limited to secondary prevention to one focusing on early reperfusion strategies.
● IV TPA (<4.5 hours)
● IA mechanical thrombectomy (<24 hours)

4. But what about the rest of the patients?
● No treatment options for patients who don’t get to the hospital late (~85% of all stroke
patients)
● Lead to stem cells emergence as a potential therapy
● Although initially designed as a replacement therapy, MSCs are not thought to replace
missing neural circuits
● Rather they seem to act as environment modifiers, altering the inflammatory landscape of
the injured brain (increasing the levels of anti-inflammatory cytokines and growth factors,
and decreasing the level of pro-inflammatory cytokines).

5. A Decade of Studies on MSCs in the Rat MCAO Model of Stroke
Journal of Cerebral Blood Flow & Metabolism (2010) 30, 653–662
Effects of administration route on migration and
distribution of neural progenitor cells transplanted
into rats with focal cerebral ischemia, an MRI study.
Lian Li1, Quan Jiang1, Guangliang Ding1, Li Zhang1, Zheng Gang Zhang1,
Qingjiang Li1, Swayamprava Panda1, Mei Lu2, James R Ewing1 and Michael
Chopp1,3
J Neurosci Res, 89(6), 833-839. 2011
Therapeutic time window and dose
response of autologous bone marrow
mononuclear cells for ischemic stroke.
Yang, B., Strong, R., Sharma, S., Brenneman, M., Mallikarjunarao,
K., Xi, X., Savitz, S. I.
Brain Res 2010 Jan 8;1306:149-58.
Therapeutic window for treatment of cortical ischemia
with bone marrow-derived cells in rats
Andréia de Vasconcelos Dos Santos1, Juliana da Costa Reis, Bruno Diaz
Paredes, Louise Moraes, Jasmin, Arthur Giraldi-Guimarães, Rosalia Mendez-
Otero
Cell Transplantation, 19, 1073–1084, 2010
Changes in Host Blood Factors and Brain Glia
Accompanying the Functional Recovery After Systemic
Administration of Bone Marrow Stem Cells in Ischemic
Stroke Rats
Ming Yang,* Xiaotao Wei,† Jing Li,† Lynn A. Heine,‡ Robert
Rosenwasser,§ and Lorraine Iacovitti*†

6. Two Current Clinical Trials
A Phase 2 Randomized, Sham-Controlled Trial of Internal Carotid
Artery Infusion of Autologous Bone Marrow–Derived ALD-401 Cells
in Patients With Recent Stable Ischemic Stroke (RECOVER-Stroke)
Sean I. Savitz, MD,Dileep Yavagal, MD,George Rappard, MD, William
Likosky, MD, Neal Rutledge, MD., Carmelo Graffagnino, MD, Yazan
Alderazi, MD., Jennifer A. Elder, PhD, Peng R. Chen, MD., Ronald F.
Budzik Jr, MD, Ronald Tarrel, DO, David Y. Huang, MD, PhD, James M.
Hinson Jr, MD On behalf of the ALD-401 Trial Group
Systemic Administration of MSCs Intracerebral Administration of MSCs

7. Overcoming the Blood Brain Barrier
●Previous work focused heavily on non-invasive routes of administration (IV, IA)
●These cells all go lungs, liver, kidney, spleen
●<1% get to brain.

8. Intravenous Injection
● IV injection proved ineffective as most
cells only made it to the spleen
● Cells that did make it to the brain were
cleared out rapidly
○ Same with other organs
Fluorescence microscopy illustrating MNCs in the brain (A), lungs (B),
spleen (C), and liver (D). Green: FITC (to label the organs
nonspecifically); red: Q-Tracker. Magnification: 3400. (E) is a histogram
showing a reduction in the number of labeled MNCs in the different
organs. The x-axis shows time after injection in hours. N 5 5 per group.
Yang, 2011

9. Intraarterial (Carotid) Injection
● Some functional affect
○ Not enough behavioral testing was done
● Rapid clearance from the brain
Brennemen, 2010

10. Blood-Brain-Barrier

11. The Goals of our Study
Intracerebrally transplant autologous MSCs in stroke patients
● Which dose is best? (1x106, 2.5x106, 5x106)
● Where do the cells go?
● How long do cells survive in the brain?
● Does infarct size change?
● How do functional recovery and behavior change?
● Do MSCs alter glial reactivity and inflammation of the brain?

12. Experimental Timeline

13. Sterile Growth Bioreactor

14. Audience Poll

15. Functional Behavior assessed through the mNSS
Group 1 (1x106 aMSCs) Group 2 (2.5x106 aMSCs) Group 3 (5x106 aMSCs)
MCAO MCAO MCAO
Txpl Txpl Txpl
Days Days Days
*P≤0.05 **P≤0.01 ***P≤0.001 ****P≤0.0001
mNSS
Score
Control (PBS injection) Experimental (MSC implant)

16. T1
T2
Day 0 Day 7 Day 30 Day 60
T1
T2
MCAO Only
(Control)
MCAO
+1mil Cells
(Group 1)
Ischemic Lesion volume and MRI Images of aMSC-Treated and Control Animals
*P≤0.05 **P≤0.01 ***P≤0.001 ****P≤0.0001
MCAO Txpl MCAO Txpl MCAO Txpl
Percent
of
Total
Infarct
Group 1 (1x106 aMSCs) Group 2 (2.5x106 aMSCs) Group 3 (5x106 aMSCs)
Control (PBS injection) Experimental (MSC implant)

17. Assessment of Cell Tracking Using Quantum Dot Nanoparticles.
Sac’d 7-Days Post-Txpl Sac’d 30-Days Post-Txpl Sac’d 60-Days Post-Txpl
INJ INJ INJ
High Power Confocal Microscopy
Group 1 (1x106 aMSCs) Group 2 (2.5x106 aMSCs) Group 3 (5x106 aMSCs)
Days Days Days

18. Immunofluorescent Staining with Ki-67 Shows the Absence of Dividing aMSCs in
the Q-dot labeled Autograft at 60-days Following Transplantation.
MSCs in Culture
MCAO-21.1
Overlay
Q-dots
Ki-67
DAPI

19. No Evidence of Transdifferentiation of MSCs into neurons or astrocytes

20. Increased Reactivity of Astrocytes and Microglia Following Stroke.
MCAO
Only
MCAO
+1X10
6
Cells
GFA
P
Contralateral
Ipsilateral
Contralateral
Ipsilateral
IBA1 Overlay
DAPI GFAP IBA1

21. Increased Reactivity of Astrocytes and Microglia Following Stroke.
MCAO
Only
MCAO
+1X10
6
Cells
GFA
P
Contralatera
l
Ipsilateral
Contralatera
l
Ipsilateral
IBA1 Overlay
DAPI GFAP IBA1

22. Corpus Callosum Volume

23. Conclusions
● Autologous MSCs (aMSCs) delivered to a stable stroke provide functional recovery and
benefit.
● aMSCs stick around for a long time but show no evidence of differentiation or
tumorigenesis.
● There was increased glial reactivity following stroke, regardless of aMSC
transplantation.
● No change to corpus callosum size, perhaps indicating less synaptic plasticity
● aMSCs are safe and effective!

24. Next Steps: Phase 1 Clinical Trial
● Currently working on Investigational
New Drug (IND) submission to the FDA
to begin human trials in Philadelphia
and Rome
● Will be the first of its kind to deliver
safe and effective autologous
mesenchymal stem cells into the brain
in chronic patients with stroke

25. Changing the game using MRI
Volume(healthy hemisphere) – Volume(infarct hemisphere) = size of infarct

26. Changing the game using MRI

27. Changing the game using MRI

28. The Aspect M7 MRI System

29. The Aspect M7 MRI System

30. Ease of Use
● Intuitive
● Plug-and-play design
● Exceptional customer service
● Open to constructive critiques

31. Q&A Session
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