Comparison of Hypothermic Storage Media

Hypothermic Storage Media Products For Cells, Tissues, and Organs

Optimizing Biopreservation Economics from Source Material to Finished Product

August 2010 © 2010 BioLife Solutions, Inc.

22!

FORWARD LOOKING STATEMENT DISCLOSURE!

2!

This presentation contains forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995 including, but not limited to, statements about BioLife Solutions, Inc. (the “Company”) and its future operating results, strategies, and product development plans. These forward-looking statements are based on current expectations and assumptions that are subject to risks and uncertainties. Actual results could differ materially from the results expressed or implied in these forward-looking statements. Factors that may cause or contribute to such differences are more fully discussed, as are other factors, in Part I, Item1A. “Risk Factors” of the Company’s Form 10-K for the fiscal year ended December 31, 2009, which is on file with the SEC and available at www.EDGAR.com. In addition, any forward-looking statements represent our estimates only as of today and should not be relied upon as representing our estimates as of any subsequent date. While the Company may elect to update forward-looking statements at some point in the future, the Company specifically disclaims any obligation to do so except as may be legally necessary, even if the Company’s estimates should change.

OTCBB: BLFS!

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!"#$$#%&!$'(')")&'!

WWe strive to be the leading provider of biopreservation tools for cells, tissues, and organs; !

to facilitate basic and applied research and commercialization of new therapies by "

maintaining the health and function of biologic source material!

and finished products during the preservation process.!

3! OTCBB: BLFS!

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!*+(,#'-!$'(')")&'!

WWe are committed to manufacturing products in accordance with!applicable current Good Manufacturing Practices and Quality System!Regulations. Every team member in the organization is responsible for!ensuring product quality during the performance of their duties and!identifying opportunities for continuous quality improvement. !

We Will:!

• Provide products and services of the highest possible standards!

to satisfy our customer needs and expectations of quality, safety,! reliability and performance!

• Encourage and promote a corporate culture of continuous !!

quality improvement!

• Maintain effective Quality Assurance Systems!

• Invest to enhance the skills of all team members!

4! OTCBB: BLFS!

•! Products:

•! Proprietary, cGMP, serum-free, protein-free biopreservation media products for cells, tissues, and organs:

•! HypoThermosol® hypothermic storage & shipping media

•! CryoStor® freeze media

•! BloodStor™ generic, cGMP cord blood stem cell freeze media

•! Services:

•! Custom biopreservation media formulation, fill, and finish into standard and custom packages

•! Customers include several competitors and the US Army

•! Contracted R & D to optimize biopreservation outcomes

•! Biopreservation media formula development

•! Verification of cell specific freezing/thawing rates and protocols

55!

PRODUCTS & SERVICES !

5! OTCBB: BLFS!

•! Regenerative Medicine is the process of engineering living, functional cell and tissue-based therapies and administering these to patients to repair or replace tissue or organ function lost due to age, disease, damage, or congenital defects

•! Target diseases include cancers, diabetes, heart disease, ALS

•! Target disorders include spinal/movement, hearing loss, vision loss, neurological (i.e., stroke)

•! Nearly all currently available and development stage regenerative medicine products and therapies utilize biopreservation processes in the acquisition of source material, isolation and manipulation of specific cells, and storage and shipment of a final product dose to a patient location

•! Biopreservation economics greatly impact product commercialization potential

•! (system costs of the shelf life & functional yield environment)

66!

REGENERATIVE MEDICINE OVERVIEW!

6! OTCBB: BLFS!

•! Market activity1:

•! 900 active clinical trials for new therapies

•! 687 companies worldwide

•! Market size and growth estimates2:

•! Market could reach $32B by 2018

•! Demand for biopreservation media3 used in process development, manufacturing, and commercial product storage, transport, and freezing will mirror the end product market growth rate

•! Increasing quality and regulatory pressure to move away from internally formulated ‘home-brew’ media and reagents

•! Limited risk of disruptive, competitive, technologies; BioLife products are the next disruptive technology that will capture share

•! Previous 9 nine quarters: BioLife product shipments to 200+ accounts

77!

REGENERATIVE MEDICINE OVERVIEW!

7! OTCBB: BLFS!

./!

.0!

.1/!

.10!

.2/!

.20!

.3/!

.30!

!" #"

45657589:;5!"5<=>=75!"98?5@!A8BC@D!E.FG!

#"2/1H!!"2//I!

./!

.1//!

.2//!

.3//!

.J//!

.0//!

!" #"

K5L97<!MB8!F=BN85O58;9:B7!"5<=9!E."G!

!"2//I! #"2/1H!

1Proteus Venture Partners, 2MedMarket Diligence, LLC, 3Management estimates

88!

BIOPRESERVATION WORKFLOW!

8!

(>PQ=O=:B7!97<!@897ONB8@9:B7!BM!OBQ8>5!

L9@58=9R!

#7SN8B>5OO!O@B8=76!B8!M855T=76!=OBR9@5<!B8!

L97=NQR9@5<!>5RRO!

UQR@Q8=76!QN!@B!9!>R=7=>9R!<BO5!

$@B8965!97<!OD=NL57@!BM!V79R!>5RR!N8B<Q>@!<BO5!

@B!N9:57@!

Enabling end to end biopreservation workflow optimization with newly integrated solutions; from source material to final product!

F=BN85O58;9:B7!WB8?XBC!=7!45657589:;5!"5<=>=75!

!""#$%&'()*+,%#,-.$)/(".%*,(',0.'&1./%&$('23,-$#/*++('23,4,5*6(7*$8,9&""68,:;.('+,

OTCBB: BLFS!

Pre-filled biologic packaging products from partners

BioLife biopreservation media products

•! Internally formulated biopreservation media products:

•! Based on historical, non-optimized formulas

•! Made daily/weekly and subject to lot-lot variability

•! Often not made with USP grade components

•! Often not made under current GMP

•! Subjected to less stringent (non-lot specific) release testing for sterility, endotoxin, mycoplasma, etc.

99!

ISSUES WITH HOME-BREW BIOPRESERVATION MEDIA!

9!

&B7SA"Y!N8B>5OO!

F9@>D!;98=9Z=R=@[!

,=L=@5<!85R59O5!@5O:76!

4\K!689<5!>BLNB757@O!

<#=*>?$*@,A="./%,

YBB858!PQ9R=@[!#75]>=57@!CB8?XBC!,=L=@5<!N85O58;9:B7!

UR=7=>9R!5^5>@O!,BC!O>9R9Z=R=@[!

&B@!BN:L=T5<!MB8!Z5O@!!N89>:>5O!

_=O@B8=>9R!MB8LQR9:B7!

OTCBB: BLFS!

•! Preservation induced cell damage and death

•! Metabolic activity decreases

•! Ionic and osmotic balances are disrupted

•! Free radicals accumulate

•! Molecular stress responses are initiated

•! Two injury mechanisms

•! Necrosis – mechanical destruction of cell membrane

•! Apoptosis – enzyme based, programmed cell death

•! Two injury exposure periods

•! During hypothermic storage and freezing

•! During re-warming/thawing

•! Delayed preservation induced cell injury – not apparent or appreciated

•! Immediate post-thaw assays do not accurately indicate cell viability

•! Perceived Viability ! True Viability

•! Traditional preservation media formulations do not adequately protect cells from preservation induced stress

•! Innovation opportunities to re-formulate preservation media to mitigate underlying causes of preservation related cell damage and death

BIOLIFE RESEARCH DISCOVERIES!

10!OTCBB: BLFS!

•! Overall system performance is constrained by limited stability, yield loss, and processing costs

•! Incremental and cumulative optimization of biopreservation processes can maximize the yield-cost relationship

•! System optimization should strive to move up and to the left

-=5R<!

$[O@5L!UBO@!

!"#$%&'()*)!

-=5R<!`!UBO@!K[79L=>!

B(*6CD:#+%,!")=(E.)#',F.$2*%,G.'2*,

BIOPRESERVATION ECONOMICS!

11!OTCBB: BLFS!

IMPACT OF LIMITED SHELF LIFE!

9%.H(6(%8,:#'+%$.('*C,-#%*').6,12!OTCBB: BLFS!

:6('(/.6,5(+%$(H&)#',G.'2*,

•! Intracellular-like media: •! Ionic concentrations are tuned to balance the intracellular state at low temperature to

reduce gradient formation and water flux

•! pH buffering capacity: •! Organic and inorganic buffers include bicarbonate and HEPES

•! Effective from 15°C to 2°C

•! Free radical scavengers

•! Osmotic stabilizing components •! To reduce and control shrinking and swelling of cells during preservation process

temperature changes

•! Serum-free energy substrates

•! Pre-formulated with cryoprotective agents (2%, 5%, or 10% DMSO)

•! No user mixing required

•! Fully-defined components

•! USP/multi-compendial or highest available grade

FORMULATION INNOVATION!

13!OTCBB: BLFS!

PROPRIETARY PRODUCTS!

HypoThermosol® !

Storage & Shipping Media !CryoStor®!

Freeze Media!

!! Formulary improvements to organ preservation media

!! Serum-free and protein-free

!! For 2 - 8°C short term storage and shipment of biologics

!! Enables functional restart upon return to normothermic temperature

!! Extends stability of specimens, pancreata, liver sections, tumor biopsies, tissue samples, bone marrow

!! Based on HypoThermosol formula

!! For cryopreservation of biologics at - 80°C to -196°C

!! Pre-formulated with 2%, 5%, or 10% DMSO

!! Serum-free and protein-free

!! Outperforms other commercial products and home-brew freeze cocktails

14!OTCBB: BLFS!

U8[B$@B8a! _[NB'D58LBOBRa!#7S_BQO5!bB8LQR9:B7!EK"$%c$58QLc"5<=9G!

$%&'()%*+,-.&/,

0&%+*'(%&&,

$%).&12'(%&&,

30$45167&8."9+-,1.:,

;<=$"=-2+>-;.+%&/,

(?@"=-8.&%"(1,&,

0.&%1,1.:A"B2/).)C12A"D"E&,,'F-8&/"G&,&-8&"H&8I26,

PRODUCT VALUE PROPOSITION!

F)$'S#&SU,($$!F#%Y4)$)4d('#%&!")K#(!Y4%K+U'$!

15!OTCBB: BLFS!

•! Preservation solutions are not devices or drugs; considered excipient or ancillary material in clinical applications

•! Ancillary material: used in manufacturing cell, gene, or tissue-engineered products, but not intended to be in final product (washed or removed) •! Excipient: component intended to be part of the final product at clinical application

•! We help customers comply with Good Tissue Practices (GTP – 1271.210) •! Highest quality reagents must be utilized in the process of developing Human Cellular, Tissue, and Tissue-Based Products (HCT/Ps)

•! “all reagents must be verified to meet specifications designed to prevent circumstances that increase the risk of the introduction, transmission, or spread of the introduction, transmission, or spread of communicable diseases”

•! Type II FDA Master Files regularly updated for both HypoThermosol® and CryoStor® product platforms

116!

REGULATORY STATUS!

16!OTCBB: BLFS!

•! Certified to: •! ISO 13485:2003 medical device quality systems management

•! Adherence to: •! 21 CFR part 820 Quality Systems Regulation of Good Manufacturing Practices for medical

devices (GMP)

•! 21 CFR parts 210 and 211 covering GMP for Aseptic Production

•! ISO 13408 manufacturing processes for aseptically-processed health care products

•! Volume 4, EU GMP Guidelines, Annex 1: Manufacture of Sterile Medicinal Products

•! ISO 14644 Clean Rooms and Associated Controlled Environments

•! Product Release Criteria: •! Sterility Tested USP <71>

•! Endotoxin Tested USP <85>

•! Cell based preservation efficacy assay

•! pH

•! Appearance

117!

QUALITY SYSTEMS!

17!OTCBB: BLFS!

•! Our biopreservation media products are incorporated into numerous development stage and commercial biologic products

•! cellular therapies

•! umbilical cord blood units

•! cells and cell lines used in drug screening

•! Critical supplier status with leading regenerative medicine and life science tools companies

118!

PRODUCT ADOPTION!

18!OTCBB: BLFS!

119!19!

EXTENDED SHELF LIFE!

alamarBlue® stain for metabolic activity

0

25

50

75

100

125 H

yp

oT

he

rmo

so

l-F

RS

FG

M (

Me

dia

)*

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

FG

M (

Me

dia

)*

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

FG

M (

Me

dia

)*

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

1 Day Storage 3 Day Storage 5 Day Storage

% R

ec

ov

ery

of

Via

ble

Ce

lls

5 Day Hypothermic Storage of Human Dermal Fibroblasts Enabled by HypoThermosol® - Assayed 24 Hours Post Storage

* Contains serum

220!20!


I$**'!!

(>:7!U[@BO?5R5@B7!

YD9RRB=<=7!Eb#'UG!

G*C!!

"=@B>DB7<8=9!(>:;9:B7!

"=@B'89>?58a!45<!

?6&*!!

&Q>R598!$@9=7!

_B5>DO@!

J/e!L967=V>9:B7!

"5<=9!C=@D!$58QL! U5RO=B8a! d=9ON97a! _[NB'D58LBOBRa!

3!K

9[!$

@B89

65!

0!K

9[!$

@B89

65!

Representative Fluorescent Micrographs Illustrating Morphology of Normal Human Dermal Fibroblasts Stored in HypoThermosol®

221!21!


0

20

40

60

80

100 H

yp

oT

he

rmo

so

l-F

RS

MS

CG

M (

Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

MS

CG

M (

Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

MS

CG

M (

Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

MS

CG

M (

Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

AQ

IX

Ce

lsio

r

Via

sp

an

1 Day Storage 3 Day Storage 5 Day Storage 7 Day Storage

% R

ec

ov

ery

of

Via

ble

Ce

lls

Extended Hypothermic Storage of Human Mesenchymal Stem Cells Enabled by HypoThermosol® - Assayed 24 Hours Post Storage

alamarBlue® stain for metabolic activity * Contains serum

222!22!


2/e!L967=V>9:B7!

&B8LBOBRaS4! YR9OL9aS,[@5! d=9ON97a! _[NB'D58LBOBRa!

1!K

9[!$

@B89

65!

3!K

9[!$

@B89

65!

Representative Fluorescent Micrographs Illustrating Morphology of Human Mesenchymal Stem Cells Stored in HypoThermosol®

I$**'!!

(>:7!U[@BO?5R5@B7!

YD9RRB=<=7!Eb#'UG!

G*C!!

"=@B>DB7<8=9!(>:;9:B7!

"=@B'89>?58a!45<!

?6&*!!

&Q>R598!$@9=7!

_B5>DO@!

223!23!



0

25

50

75

100

125 H

yp

oT

he

rmo

so

l-F

RS

Me

dia

*

No

rmo

so

l-R

Pla

sm

a-L

yte

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

Me

dia

*

No

rmo

so

l-R

Pla

sm

a-L

yte

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

Me

dia

*

No

rmo

so

l-R

Pla

sm

a-L

yte

Ce

lsio

r

Via

sp

an


% R

ec

ov

ery

of

Via

ble

Ce

lls

Recovery of Human Osteoblast Cells Following Hypothermic Storage Assayed 24 Hours Post Storage

- Differentiated Following 5 Day Hypothermic Storage of Undifferentiated hMSC in HypoThermosol

224!24!

POST-PRESERVATION DIFFERENTIATION CAPACITY!

2/e!L967=V>9:B7!

&B8LBOBRS4! YR9OL9S,[@5! d=9ON97! _[NB'D58LBOBR!

3!K

9[!$

@B89

65!

f!K

9[!$

@B89

65!

Representative Fluorescent Micrographs Illustrating Morphology of Human Osteoblast Cells Following Hypothermic Storage

- Differentiated Following 5 Day Hypothermic Storage of Undifferentiated hMSC in HypoThermosol

I$**'!!

(>:7!U[@BO?5R5@B7!

YD9RRB=<=7!Eb#'UG!

G*C!!

"=@B>DB7<8=9!(>:;9:B7!

"=@B'89>?58a!45<!

?6&*!!

&Q>R598!$@9=7!

_B5>DO@!

225!25!


0

25

50

75

100

125 H

yp

oT

he

rmo

so

l-F

RS

RP

MI

(Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

RP

MI

(Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

Ce

lsio

r

Via

sp

an

Hy

po

Th

erm

os

ol-

FR

S

RP

MI

(Me

dia

) *

No

rmo

so

l-R

Pla

sm

a-L

yte

Ce

lsio

r

Via

sp

an


% R

ec

ov

ery

of

Via

ble

Ce

lls

24 Hour Post Storage Recovery of Human Dental Pulp Stem Cells Following Hypothermic Storage

Commercial Solution Comparison: Serum and Serum-Free Media


•! Scientific validation across a broad set of biologics, offering significant performance and quality advantages

•! Proven ability to extend stability and improve post-preservation viability, recovery, and function of specimens, source material, and isolated cells

•! Viewed as a critical manufacturing reagent supplier by numerous cell therapy companies and cell suppliers

•! Demand for our proprietary products will closely follow pace of increased regulatory and quality oversight of reagents used in clinical product development, processing, storage, transportation, and infusion

226!

TECHNOLOGY SUMMARY!

26!OTCBB: BLFS!

Technology

Comparison of Hypothermic Storage Media