Soterias Medical

SOTERIAS MEDICAL

A Novel Stem Cell Delivery Device

Michael Clark, Prateek Gowda, Angelica Herrera, Seung Jung,

Michael Mow, Arianne Papa, Annabeth Rodriguez, Jose Solis

Project Advisors: Luis Garza, M.D., Ph.D (Johns Hopkins Hospital)

Sewon Kang, M.D. (Johns Hopkins Hospital)

Robert Allen, Ph.D. (Johns Hopkins University)

Johns Hopkins University

STEM CELL MARKET

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● Global stem cell market is

growing at 13.6% from $5.6

billion in 2013

● Currently, 4681 clinical trials on

stem cell therapies in the US

(3000 in Phase I or II)

● Skin Stem Cell Therapies

STANDARD OF CARE

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STANDARD OF CARE

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Traditional Needle Injection

● No consistency in▪ Thaw rate

▪ Injection volume

▪ Injection rate

▪ Positional control

● Risk of infection

These factors lead to a reduction in cell viability.

CLINICAL NEED

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There is a need for a device that allows

physicians to deliver stem cells to target

dermal regions at adjustable volumes with

minimal risk of infection or viability loss.

SOTERIAS MEDICAL

Factor Standard of Care Replicel Soterias Medical

Controlled Injection Rate ☓ ✓ ✓

Minimal Risk of Infection ☓ ☓ ✓

Built-in Thawing Capability ☓ ☓ ✓

Adjustable Volume ✓ ✓ ✓

Consistent Angle of Injection ☓ ✓ ✓

PRODUCT COMPARISON

PRIOR DESIGNS

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CURRENT DESIGN

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DERMAL INJECTOR PROCEDURE

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BASE STATION

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Features

● Docking station for

device when not in use

● Charging station

● LCD screen

● Adjust injection settings

○ Cell thawing

○ Injection rate

○ Injection volume

Features

● Controlled feedback

system heats pads to

37°C

● Consistent and quick

thaw time ensures cell

viability

● Eliminates need for water

bath in procedure room

HEATING ELEMENT

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Features

● Larger injection volume

results in fewer number of

injections.

● Closed-loop system

reduces contamination.

● Luer lock connections.

● Microliter precision from

tubing

CLOSED - LOOP SYSTEM

SOTERIAS MEDICAL

Features● Peristaltic Pump

● Motor

● Arduino

○ Motor Control

○ Temperature negative

feedback loop

● Raspberry Pi

○ Program the device

○ User-friendly interface

for physicians

ELECTRONICS

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Features

● Septum improves device

safety

● Angle guards provide

positional control

● Easy setup and safe

removal of disposables.

OTHER FEATURES

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TESTING AND EVALUATION

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Factor Test Satisfactory Result

Thaw Time Thaw dead cell slurry -196℃→ 37℃, < 2-3 minutes

for 1.0 mL

Cell Viability Ex vivo: Injection Rates and

Diameters

In vivo: Clinical Trial

<40% Reduction in Viability

Injection Volume Determine density of dead cell

slurry, measure mass of

injected volume

Accuracy of ±1 μL

Human Factors Qualitative Feedback No more difficult than current

standard of care

1. Mouse spleen cells to

mimic fibroblasts

a. Used Countess

Automated Cell Counter.

a. Unable to differentiate

between RBCs and

mouse spleen cells

a. Needed ACK RBC lysis

buffer

TESTING RESULTS

TESTING RESULTS

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2. Sole fibroblasts from Dr.

Garza’s lab

a. Inconsistent cell density

when pipetting onto

hemocytometer slides

a. All cells appeared viable

immediately after injection

TESTING RESULTS

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3. L929 mouse fibroblasts from

Dr. Logsdon

a. 60,000/mL cell density too low

to see increased shearing

forces

a. All cells appeared viable

immediately after injection

a. Trypan blue had debris making

images unclear.

TESTING RESULTS

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4. Thermocouple Negative

Feedback Loop

a. Stabilize temperature at

37.0°C with ±0.5°C variation

a. Reduced error with more

frequent temperature checks

PROTOTYPE BUDGET ANALYSIS

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Handheld Component Cost for Device

Plastic PLA Shell $0.16

Buttons $0.16

LEDs $1.32

Pins, Springs, Hinges $13.34

Pump Components $26.55

Heating Components $67.57

Total $109.10

Base Station Component Cost for Device

Plastic PLA Shell $0.18

Buttons $0.90

Power Cord, Contact Leads $9.65

LCD Screen $35.00

Total $45.73

Disposable Component Cost

23 Gauge Needle $0.18

Tubing, Luer Lock $1.42

Cryogenic Bag $31.00

Total $33.25

ACCOMPLISHMENTS & AWARDS

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ACCOMPLISHMENTS

Base station

Prototype angle guards

Pumping mechanism

Heating mechanism

Cell testing

Heating testing

Closed-loop system

Handheld shape design

AWARDS

First Place, Carnegie Mellon University McGinnis

Venture Competition

Finalists, TigerLaunch

Finalists, The Big Sell

Finalists, Towson Incubator

Finalists, Carnegie Mellon Venture Competition

Semifinalists, ASME iShow

Semifinalists, JHU Business Plan

Semifinalists, Tulane Business Model

✓

✓

✓

✓

✓

✓

✓

✓

SOTERIAS MEDICAL

SHORT TERM

Visualize Injection Progress

Angle Guards

Shorter Needles

Reduce Size with a New Pump Design

Cartridge

User Interface development

Testing

Power Contacts

LONG TERM

Explore FDA Pathways

Provisional Patent

Intellectual Property

Business Plan Competitions

Partnering

Team Expansion

Exit Strategy

NEXT STEPS

Dr. Robert Allen Dr. Elizabeth Logsdon Dr. Warren Grayson Dr. Zijun Zhang

ACKNOWLEDGEMENTS

SOTERIAS MEDICAL

Dr. Luis Garza

Dr. Youseph YazdiDr. Soumyadipta

Acharya

Dr. Hai-Quan Mao

Mihika ReddyAaron ChangNathaniel Leon

Dr. Sewon Kang

Amadeus Zhu Tabetha Ratliff

Appendix Index● Markup Price

● 5 Year Startup Expenses

● Marketing

● Distribution

● Sales Channels

● Client Base

● 10 Year Financial Breakdown

● Cryomaterials

● Cryobags

● Cryotubes

● Cell Viability Post Injection

● Peristaltic Pumps

● Tubing Dead Space

● Needle Dimensions: Length

● Needle Dimensions: Angle

● Dermal Fibroblast Size

● Anatomy of the Dermis

● Epidermal Thickness I

● Epidermal Thickness II

● Replicel RCI-01, RCI-02

● Testing:

○ Injection Volume

○ Human Factors

○ Thaw Time

○ Injection Depth

○ Cell Viability

○ Patient Discomfort

○ Delivery Time

● Detailed Injection Procedure

● Closed-Loop System Details

● Electrical Components

● Motor

● Springs, Clips, Hinges

● Heating

● Pump

MARKUP PRICE

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Markup Price

Device $199.99

Disposables $ 59.99

Manufacturing Cost

Device $154.83

Disposables $ 33.25

Total: $188.08

5 Year Startup Expenses

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MARKETING

SOTERIAS MEDICAL

Market through interactions with hospital sales representatives

Foster awareness through scientific community

Advertised at medical conferences and online

Estimated 9,600 dermatologists and 7,800 dermatology practices in the U.S.

More than 91.5 million people qualify for treatment

Secondary Target:

Universities and other

research institutions.

DISTRIBUTION

SOTERIAS MEDICAL

Primary Target:

U.S. hospitals/clinics with a

commitment to the cutting edge

of medical technology.

SALES CHANNELS

SOTERIAS MEDICAL

PHASE I - DIRECT

● Seek out leaders in stem cell research

● Target physicians most likely to adopt

novel devices

● Hire and utilize in-house marketing

experts

● Engage customers with face-to-face

encounters

PHASE II - INDIRECT

● Market the device to medical supply

distributors to drive continuing sales

● Pitch the device nationwide to hospitals

and hospital representatives

● Engage private practices throughout the

country

● Others approach our company

CLIENT BASE

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Trusting relationship

Customer loyalty

Customer trust

Improved outcomes

Success of product

Technical assistance

10 Year Financial Breakdown

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Cryomaterials

● Cryogenic materials have special

properties that make them particularly

hazardous to use in the solid, liquid or

gaseous states.

● Cryogenic materials are characterized by

severe low temperature (-60℃ to -270℃).

● Most cryogenic liquids are odorless and

tasteless when vaporized; however, when

exposed to the atmosphere they create a

highly visible dense fog that dissipates

with warming.

● Cryogenic temperatures are achieved by

liquefaction of gases, most commonly

helium, hydrogen, nitrogen, argon, oxygen

or methane.

Cryobags

● Designed to process, grow, and

store cells and tissues from -196°C

to over 200°C

● Bags come in various sizes

(Freeze volume range: ~10-200mL)

● Made of two materials

o Ethyl Vinyl Acetate (EVA)

o Fluorinated Ethylene Propylene

(FEP)

● Four known suppliers:

o Charter Medical

o Instant Systems

o OriGen Biomedical

o Quest Biomedical

Cryotubes

● Designed for storing biological

material at temperatures as

low as -196°C

● Tubes come in various sizes

(Freeze Volume Range: ~0.5-

5mL)

● Major Suppliers:

○ Sigma-Aldrich

○ Corning

○ ThermoScientific

Cell Viability Post-Injection

Peristaltic Pumps

Pros Cons

Sterile pathway Tubing Dead Space

Good for shear-sensitive materials Pulsating output at low flow rates

Very fine volume/rate control Need to “calibrate”

Tubing Dead Space

Source: http://www.had2know.com/technology/curved-pipe-volume.html

● Tube volume - including the exact

volume of any bends - can be

mathematically determined

● Rough estimate based on the fill

volume of appropriate lengths of tubes

were obtained

o 76uL

Needle Dimensions: Tube

Source: Sigma Aldrich

Needle Dimensions: Angle

Source: Sigma Aldrich

≈9°

≈23°

≈37°

αO.D.

O.D./sin(α)

Dermal Fibroblast Size

About 20μm diameter

Anatomy of the Dermis

Epidermal Thickness

Up to 370μm thickness (fingertip)

Epidermal Thickness

About 84μm average thickness

2003

Total Skin Thickness

Replicel

Testing: Injection Volume

● Objective: Device should be able

to deliver desired volume (5μL -

1000μL, 5μL increments) with

accuracy of ± 1μL

● Detailed Experiment:o Pseudo-calibrate pipette using

deionized water

o Determine dead-cell slurry density

o Determine injection volume

accuracy using skin sample

Testing: Human Factors

● Objective: Device should not be any

more difficult than current standard of

care

● Detailed Experiment:o Present prototype to physician

o Observe handling of device and record any

comments or questions he/she brings up

o Instruct physician on proper use of device

o Observe handling issues and/or cumbersome

features

o Acquire feedback on: ambidexterity,

complexity, size, steadiness, weight

Testing: Thaw Time

● Objective: Device should thaw solution at the

same rate as a 37°C water bath or human hand

● Detailed Experiment:o Cryobag/cryovial is frozen in liquid nitrogen

o Samples are removed from liquid nitrogen and thawed

via 3 different methods:

▪ 37°C water bath

▪ Hand heat

▪ Injector device heating element

o Amount of time for entire solution to liquefy will be

measured

Testing: Injection Depth

● Objective: Deliver stem cells at a specific vertical depth at a ±

10% depth accuracy

● Detailed Experiment:o Ex Vivo:

▪ Device is set to predetermined depth and volume

▪ Dead stem cells dyed with trypan blue and loaded into device

▪ After injection, stem cells will be sent for histology

▪ Images quantified by measuring distance from top of epidermis

to top and bottom of cell bolus

o In Vivo:▪ Device is set to predetermined depth and volume

▪ Stem cells will be injected into patient

▪ OCT/ultrasound imaging will be used to image testing site of

patient immediately after injection and after a set time to allow

for diffusion

▪ Images quantified by measuring distance from top of epidermis

to top and bottom of cell bolus and area of cell bolus

Testing: Cell Viability

● Objective: Loss in viability due to injection should be no greater than current

standard of care, preferably < 40% reduction in viability

● Detailed Experiment:

o Ex Vivo▪ Harvest cells from mouse spleen

▪ Load cells into device with cryosolution

▪ Inject cells onto a petri dish using device as well as a needle and

syringe using different rates of injection and needle diameters

▪ Cells will be then stained using live/dead assay

▪ Stained cells on petri dish can then be counted under a light

microscope using a cell counting software to determine cell viability

o In Vivo▪ Two 700 µL injections will be performed on lower back of patient:

one with device, one using needle and syringe

▪ Three months later, cell viability will be qualitatively evaluated

▪ Cell viability for each injection technique will be evaluated using

difference in area of growth and OCT/ultrasound/staining methods

Testing: Cell Viability

Control: 100 uL Pipette 100 uL Needle and Syringe 100 uL Pump

Testing: Patient Discomfort

● Objective: Device should not pose

discomfort greater than that of the

current standard of care

● Detailed Experiment:o Inject saline solution at desired depth

and volume for normal procedure into

fully informed volunteer human test

subject using prototype

o Inject into upper buttocks of test subject

o Record duration of procedure

o Survey test subjects on degree of pain

Testing: Delivery Time

● Objective: Device should be able to thaw

1 mL solution in approx. 2 minutes and

deliver the amount in approx. 1 minute

● Detailed Experiment:o Thaw solution in cryostore bag

o Load bag into prototype

o Adjust volume and depth dials to desired

values

o Inject cells using prototype into upper buttocks

of patient

o Record total time and any complications

Detailed Procedure

1. Lab technician freezes cells in cryobag at -196℃. This is all performed in a hood.

2. “Runner” brings cells from freezer to clinic where procedure will take place while cells are on dry ice.

3. Physician receives cells, feeds cryobag, tubing, and needle into device while sitting in base station.

a. Clip into device

b. Clip tube in

c. Feed tube through pump

d. Clip needle

4. Close device

5. Turn on/program base station (desired injection rate, volume, thawing temperature)

6. Let cells thaw. Device will stop heating when bag is thawed and “beep.”

7. Remove from base station.

8. Pick angle guard/install on tip of device. (optional)

9. Remove cap from needle

10. Remove dead space in tubing by pushing button on device to eject air.

11. Prep for injection

12. Inject

13. Open device, remove everything via bag

14. Place device back in base station

Closed - Loop SystemCryobag

● Holds 7-12mL volume

● 3.0 cm x 6.3 cm

Luer Lock ● Male Luer with Spin Lock to Barb

● Fits 1/32 inch ID Tubing

Tubing● High-Purity Silicone Rubber for

Peristaltic Pump

● OD: 3/32" ID: 1/32"

● 1/32" Wall Thickness

● Sub-microliter precision

Needle● 23 gauge

Electrical Components

The device utilizes an Arduino Micro Board in

the handheld injector and a Raspberry Pi in the

base station

Arduino Micro Functions:

● Motor Control

● Temperature Feedback on Heating

System

● Injection Command

● Internal Power Supply 12V

Raspberry Pi Functions:

● User-friendly color touch screen

● Serves as interactive medium for

dermatologist to set injection volume,

speed and cell temperature

Motor Specifications

12V 1/64 Geared Stepper Motor

1024 Step / Revolution Resolution

100 N/mm Holding Torque

Weight: 37g

Diameter: 28mm

Height: 20mm

Provides high resolution and controlled

movement without the need of an encoder

Springs, Hinges, and Clips

Springs

- Located behind heating pads.

- Controls the force of contact

between device and cyro-bag.

Hinges

- Located on the side of the device.

- Integrated with design.

Tube Clip

- Located towards the head of the

device.

- 3D printed onto device.

- Holds tubing in place during

injection.

Heating

Heating pads

- 2 5V pads

- Squeeze bag together using springs to

heat evenly

Relay

- Electrical component used to switch pads

on/off when the temp. reaches 37℃Arduino Uno

- Used to receive feedback from heating

pads and control relay

Thermocouple

- Temperature probe that measures temp.

of bag/pads during thawing

Power

- 9V DC power supply from wall plug

Pump Specifications

3 cylinder peristaltic pump

Currently PLA but will eventually

be made out of aluminum or

stainless steel for durability.

Setup provides 1uL precision.