Cryopreservation for Cell Therapy CoolCell® SV2 and SV10 packages

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May 2014

•  Equal or better results than programmable step-down

freezers and isopropanol-based freezing containers •  Simply fill with vials and place -80oC freezer •  Radially-symmetric thermal exchange design ensures

uniform cooling profiles to all vial positions •  Composed of high-density materials used in aerospace

industry for lightweight & insulative properties •  Highly thermo-conductive alloy core •  Models include conduction and/or micro-convection air

flow control

Passive controlled-rate cell freezing

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CoolCell® controlled-rate freezing containers*

CoolCell® containers deliver equal or better results than programmable freezers and isopropanol-based freezing containers.

*Patent pending.

Five consecutive freezing runs show extremely consistent thermal profiles.

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CoolCell® containers ensure reproducible freezing profiles

Stem Cells and Cell Therapy Products •  Testing and data performed by USCF (Jeff Bluestone lab), Roslin Cellab,

TxCell SA and others Cell Lines

•  LnCap, HTB77, A549, others •  Data and testing done by Lonza (Germany)

Primary Cells •  Data and testing done by labs at UCSF and ZenBio

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CoolCell® containers are compatible with many cell types

See CoolCell® freezing container performance data here: http://www.biocision.com/resources/coolcell-performance-data

Post-thaw viability and cell yield of T-regs frozen in controlled-rate freezer and CoolCell® container Human T-regs aliquots were transferred into closed-system 2ml glass ampules in a cryopreservative at 1 to 10 x 106 cells/mL and then frozen in using the optimal -1C/min freeze rate. Post-thaw cell viability and yield were calculated using a flow cytometer. Post-thaw cell viability was 91.7 ± 3.7% from the CoolCell device and 91.7 ± 4.0% from the programmable freezer. The cell yield was also comparable, leading TxCell to convert from programmable controlled-rate freezer to CoolCell containers for Phase 2b clinical trials.

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Human T-regs for Cell Therapy Data provided by TxCell

Post-thaw viability of RC-10 after 4 freezing methods Human embryonic stem cells, RC-10, grown to confluence were harvested and suspended in a cryopreservative at 2 x 106 cells/vials and then frozen in 1.0 ml cryogenic vials using the methods above. After at least two weeks in LN2, 3 vials from each treatment were thawed and cells were counted immediately (Day 1) or after three days growth (Day 3). The CoolCell container freezing method led to a slight increase in the number of viable cells on day 1 and a 33% increase in the number of cells after 3 days of growth.

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Human Embryonic Stem Cell (RC-10) Data provided by Roslin Cellab

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Cell proliferation after 4 freezing methods Proliferation analysis of the RC10 cells was performed using the Alamar Blue reduction assay. Assays were performed in 96 well plates seeded at 10,000 cells per well at 6, 24 and 30 hours. (Data is not shown for 30 hours, but the same trend was observed.) Cells frozen in the CoolCell container grow more quickly leading to higher proliferation rates and more cells.

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Human Embryonic Stem Cell (RC-10) Data provided by Roslin Cellab

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Cell adherence after 4 freezing methods Images of human embryonic stem cell line RC10 at day 1 post resuscitation from different freezing methods at 10X magnification using three different freezing methods. Differentiation was measured by flow cytometry assessment of both pluripotency status (RC-7 p18, RC-10 p27) and differentiation markers (RC-7 p19-20, RC-10 p28-29). Cells frozen in the CoolCell adhere to the plates and grow more quickly, while maintaining undifferentiated state.

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Human Embryonic Stem Cell (RC-10) Data provided by Roslin Cellab

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CoolCell Styrofoam Box

Paper Towels

“Mr. Frosty”

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CoolCell® controlled-rate freezing containers

CoolCell® freezing containers accommodate a variety of sample vessels: •  2 mL to 5 mL cryogenic vials •  2 mL injectable ampules •  10 mL injectable ampules

CoolCell® SV2 and CoolCell® SV10 freezing containers for ampules •  Reproducible -1°C/min freeze rate •  Replaces programmable freezers and isopropanol-

based freezing containers •  For 2 mL or 10 mL injectable ampules

CoolCell® Stem Cell Cryopreservation Systems

•  Includes CoolCell® freezing container and ice-free cold cell preparation system

•  Ideal for use in the hood •  Ice-free cooling system is easy to use, compact, and

keeps all vials uniformly cold (0.5 to 4.0°C) for up to 4 hours

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CoolCell® Stem Cell Cryopreservation Systems

CoolCell® SV2 Stem Cell Cryopreservation System

CoolCell® SV10 Stem Cell Cryopreservation System

CoolCell® SV2 for 12 x 2 mL injectable ampules

CoolCell® SV10 for 6 x 10 mL injectable ampules

BioT® ULT Transporter and Workstation •  Dry ice-based systems utilize patented

DIR™ technology to ensure ultra-low cooling (-80°C to -50°C) for hours while open

•  Closed-cell high density polyethylene foam exterior is highly insulative and not cold to the touch

•  Vertical chamber space allows ample ultra-low temperature work zone

•  Superior performance compared to transporting samples directly on top of dry ice

•  Available in a variety of sizes for portable transport, stationery processing or cart-based mobility

•  Internal: 11.2 x 6.5 x 9.0 in / 28.4 x 16.5 x 22.8 cm

•  External: 18.0 x 11.2 x 12.5 in / 45.7 x 28.4 x 31.8 cm

•  Holds up to 8 standard cryostorage boxes or 2 CoolCell freezing containers

BioT® ULT Mobile Workstation •  Portable dry-ice based cooling system •  Patented DIR™ technology ensures entire

chamber is in ultra-low temperature range (-80°C to -50°C) for over 8 hours

•  Ideal for transporting frozen samples or for use as a compact freezer

•  Internal: 11.2 x 6.5 x 9.0 in / 28.4 x 16.5 x 22.8 cm

•  External: 18.0 x 11.2 x 12.5 in / 45.7 x 28.4 x 31.8 cm

•  Holds up to 8 standard cryostorage boxes or 2 CoolCell freezing containers

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BioT® Temperature Stability Systems

BioT® ULT Transporter. Over 8 hours of <-50°C cooling. Compact with carrying handles. Ideal for transport or as a compact, portable freezer

BioT® ULT Workstation. Over 15 hours of <-50°C cooling in approx 10” vertical work zone.

"As we advance our promising clinical research program for Ovasave® to a Phase IIb trial, the BioCision product portfolio and technology platform present the ideal range of capabilities and quality standards necessary to help us maintain the essential levels of standardization and control we need to be successful."

Arnaud Foussat, Vice President Research and New Products, TxCell

“This is by far and away the best benchtop technology to enter the field of cell cryopreservation for decades.”

John Gardner, Roslin Cellab

“We run a registry, in which large amounts of PBMCs are processed for long-term cyropreservation.………Overall, the CoolCell has proven to us to set a new bar in cryopreservation.”

Rohit G., Stanford University

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Customer Testimonials

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