“derivation and uses of highly pure and scalable lineage-specific stem cell … · 2020-01-26 ·...

“Derivation and uses of highly pure and scalable lineage-specific stem cell lines and their cell products”

January 23, 2020

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Forward Looking Statements

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The matters discussed in this presentation include forward looking statements which are subject to various risks,uncertainties, and other factors that could cause actual results to differ materially from the results anticipated. Such risks anduncertainties include but are not limited to the success of AgeX Therapeutics in developing new stem cell products andtechnologies; results of clinical trials of such products; the ability of AgeX and its licensees to obtain additional FDA andforeign regulatory approval to market products; competition from products manufactured and sold or being developed byother companies; the price of and demand for such products; and the ability of AgeX to raise the capital needed to finance itscurrent and planned operations. Any statements that are not historical fact (including, but not limited to statements thatcontain words such as "will," "believes," "plans," "anticipates," "expects," "estimates") should also be considered to beforward-looking statements. Forward-looking statements involve risks and uncertainties, including, without limitation, risksinherent in the development and/or commercialization of potential products, uncertainty in the results of clinical trials orregulatory approvals, need and ability to obtain future capital, and maintenance of intellectual property rights. As actualresults may differ materially from the results anticipated in these forward-looking statements they should be evaluatedtogether with the many uncertainties that affect the business of AgeX and its subsidiaries, particularly those mentioned in thecautionary statements found in AgeXSecurities and Exchange Commission filings. AgeX disclaims any intent or obligation toupdate these forward-looking statements.

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The PureStem Embryonic

Progenitor Cell Bank

PureStem®: Industrial Scale Source of Young Cells/Cell Products

•Scalable•Pure•Stable Identity•Diverse•Young/Highly Regenerative

Addresses cell manufacturing bottleneck to treat large aging patient population

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PureStem Technology

Off-the-Shelf Therapeutics

UniverCyte: HLA-G+ PureStem Lines

Cell Products - ExosomesCell Therapy

PureStem®: Industrial Scale Source of Young Cells/Cell Products

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Regenerative Capacity is Highest before EFT (Embryonic to Fetal Transition)

Evolution

Deve

lopm

ent a

nd A

ging Weismann Barrier

Pluripotency

Embryo

Fetus

Newborn

Evolution

Adult

EFT

PT

NT

AT

Development recapitulates loss of immortal regeneration during evolution

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CellsExosomes

Aging Adult

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Therapeutic Angiogenesis

EmbryonicProgenitors

Pluripotent stem cells

iTR: induced Tissue Regeneration

Can We Induce Regeneration using Embryonic Progenitors?

Highly Regenerative Progenitors

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Pluripotent Stem Cells

Scaling at the Embryonic Progenitor StagePu

reSt

emTe

chno

logy

>200-fold diversityScalable, clonallypurified progenitors

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Pluripotent Stem Cells

Clonal Selection

Condition 1

1 2 3 4 5 6 7 8Selection for Growth Medium

Candidate Cultures

Partial Differentiation

Condition 2 Condition 3 Condition 4 Condition 5 Condition 6…

* * *

Expansion and Banking

PureStem 1 PureStem 3…PureStem 2

Industrial Scaling of Young Progenitor Stem Cells

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Cell Identity + Diversity: >200 Distinct Cell TypesMicroarray analysis

BoneCartilage

Smooth Muscle

Endothelial

Pericyte

Brown Fat

Fate Space Mapping

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Clonal Osteochondral Progenitors – Orthopedics

OTX-CP03

Experimentally-induced trauma

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Clonal Brown Fat Progenitors – Obesity

UCP1

3D Culture in HyStem®

Oil Red O

2D Culture

Phase

1212

E69

T42

Precise Neural Crest Cell Types - CNS

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Clonal Smooth Muscle Progenitors - Cardiovascular

0

2000

4000

6000

8000

10000

12000

14000RF

U

Differentiation Conditions

MYH11

**

14

Clonal Endothelial Progenitors - Cardiovascular

Monoclonal Endothelium

99.7%

Endothelial Surface Markers

VEGF Induced Angiogenesis

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PureStem ECAdult ECNon-EC Lines

EC-Related Genes

PureStem Endothelial Embryonic Progenitor Cell Lines

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Retention of Embryonic Phenotype in PureStem Embryonic Progenitors

PureStem ECAdult EC

PureStem EC

Adult EC

HUVEC

HCMEC

Potential to Produce Highly Regenerative Cell Products

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Population Doublings

22 34 43 55 65 72 82 90

Dec

reas

ing

Telo

mer

e Le

ngth

with

Age

PureStem Cell Lines have Long Telomeres

TRF Length(kbp)

15

10

5

Age

Neonatal Hayflick Limit

Germ-Line & Pluripotent SCs(Telomerase +)

Somatic(Telomerase -)

Young Cells = Long Telomeres

18Regen Med 2012 Jul;7(4):481-501

Very Long Replicative Lifespan

Embryonic Telomere Length = High Scalability

Adult MSCs

Popu

latio

n Do

ublin

g

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Industrial Scale Cell ManufacturingConsistent Large Scale Cell/Exosome Production from Single Cell Source

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Opportunity: PureStem® Technology applied to Exosome Therapeutics

+ ve Ctrl - ve CtrlPureStem Exos

PureStem Exosomes Lack of MHC Antigens

• Safety • Simplicity• Potency• Delivery/Dosing• Convenience/Storage• Lower immune risk

• Off-the-Shelf Product• Faster Track to Clinic

Leveraging Exosome Therapeutics

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Pre-EFT (Embryonic to Fetal Transition)

Exosomes

Cellular Network

Exosomes: Secreted Vesicles for Intercellular Communication

RNAProteinLipids

Highly RegenerativeExosome Cargo

PureStem Cell Library


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Selection of PureStem Exosomes in Wound Healing Assay

0h

14h

Exo-Free

Exo-Free 30-MV2-14

30-MV2-14 VEGF

VEGF

VEGF 10

RPI-MV2-8

30-MV2-14

Exo-Free

20

40

60

80

100

0

Rela

tive

Wou

nd D

ensit

y (%

)

Time (Hours)6 12 18 240

Exo-Free

Positive ++

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0h

14h

Exo-Free

Exo-Free

VEGF

VEGFExo-Free

VEGF 10

30MV2-19

30-MV2-10

0

Time (Hours)6 12 18 240

20

40

60

80

100

Rela

tive

Wou

nd D

ensit

y (%

)

Exo-Free

VEGF 2

Positive +

30-MV2-19

30-MV2-19

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20

40

60

80

100

Rela

tive

Wou

nd D

ensit

y (%

)

VEGF

MSCE69

Exo-Free

06 12 18 240

Time (Hours)

0h

14h

Exo-Free

Exo-Free Primary MSC

Primary MSC VEGF

VEGFExo-Free

Negative -

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Selection of Angiogenic PureStem Exosomes

Need graph showing Exo Free and VEGF

Tube length vs time

AUC? VEGF +ve ControlExo-Free –ve Control

Net

wor

k Le

ngth

(mm

/mm

2 )

Time (hours)

Exo-Free

VEGF

ControlsPlate Cells

Evaluate Network

Co-Culture Primary Fibroblasts + Endothelial Cells (HUVEC)

Add Exos

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VEGF +ve ControlExo-Free –ve ControlExo-Free

30-MV2-14

VEGF

RP1-MV2-8

Positive +

Net

wor

k Le

ngth

(mm

/mm

2 )

Time (hours)

Plate CellsEvaluate Network


Add Exos

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VEGF +ve ControlExo-Free –ve Control

Negative -

**

VEGF

PCMW10

Exo-Freens

Net

wor

k Le

ngth

(mm

/mm

2 )

Time (hours)

Plate CellsEvaluate Network


Add Exos

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Plate Cells

VEGF +ve ControlExo-Free –ve Control

Evaluate NetworkAdd Exos

Negative -

**

VEGF

MV2-19

MV2-10

Exo-Free

ns

HUVEC

Net

wor

k Le

ngth

(mm

/mm

2 )

Time (hours)


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Source Cell Line Cell Type Wound Healing

Angiogenesis

30-MV2-10 Endothelial + -30-MV2-14 Endothelial ++ +30-MV2-19 Endothelial + -RP1-MV2-8 Pericyte ++ +

PC-M Pericyte - -W10 Smooth Muscle - -E69 Neural Crest - -

MSC-WJ MSC - -HUVEC Endothelial nd -

Strong Wound Healing Correlates with Angiogenic Activity


PureStem(Embryonic)

Source Cell Line Cell Type Wound Healing

Angiogenesis

30-MV2-10 Endothelial + -30-MV2-14 Endothelial ++ +30-MV2-19 Endothelial + -RP1-MV2-8 Pericyte ++ +

PC-M Pericyte - -W10 Smooth Muscle - -E69 Neural Crest - -

MSC-WJ MSC - -HUVEC Endothelial nd -

Primary (Adult)

PureStem(Embryonic)

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Group 130MV2-14

Group 2RP1-MV2-8

Diversity: Exploring Progenitor Space

Group 3Group 4

PureStem Transcriptomics

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Diversity: PureStem Exosome RNA Cargo Content

Angiogenic microRN

As

Angiogenic mRN

As

Production Cell Lines Production Cell Lines

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Scalability: Stable Production of Embryonic Progenitor ExosomesEarly Vs Late Passage

T-Flask Vs BioReactor

0

10

20

30

40

50

60

70

80

90

0 20 40 60 80 100

Popu

latio

n D

oubl

ing

Time (Days)

Primary-MSC

PureStem Endothelial

High Production Capacity

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Exosome Supercharging: Loading RNA Cargo into Exosomes

100

80

60

40

20

0 5 10 15 20

Exo-Free

Scramble

miR-126

AngiomiR-126 increases Exosome Wound Healing Activity

0Re

lativ

e W

ound

Den

sity

(%)

Time (Hours)

Inactive Exosomes

Charge with RNA

ActiveExosomes

Engineering PureStem Exosomes

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• Angiogenesis

• Senolytic Activity• Epigenetic Rejuvenation

Summary: Applications of PureStem Cells and Cell Products (Exosomes)

PureStem Cell Library

Exo- iTR


Partial Reprogramming

Natural and Engineered Exosomes

Heart Disease

Stroke

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Acknowledgements

• Jieun Lee• Wei Guo• Michael West• Hal Sternberg

“We invite you to partner with us.”

“derivation and uses of highly pure and scalable lineage-specific stem cell … · 2020-01-26 ·...

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