ThawSTAR™ Cell Transport and Thawing System is a first-of-its-kind technology platform that combines ultra-low temperature vial transport and automated cell thawing in a convenient and compact format. The system replaces manual methods of cell thawing such as swirling frozen vials in communal water baths and warming vials between hands - methods that are not standardized and potentially jeopardize the integrity and safety of the sample - and enables thawing in a laminar flow hood. The complete system is comprised of a ThawSTAR™ CFT Transporter and a ThawSTAR Automated Cell Thawing System. Together, they provide better cell recovery and viability and less variability over conventional methods.

The CFT Transporter is a portable, dry ice-based module that enables ultra-low temperature (~75°C) transport of frozen vials from the storage location to the thawing location. The transporter keeps up to 5 vials at ultra-low temperature for up to one hour with a small amount of dry ice and ensures that all vials are at the same temperature when inserted into the thawing instrument.

The ThawSTAR automated cell thawing instrument utilizes patent-pending STAR™ sensing technology that employs multiple detection mechanisms to determine vial temperature, phase change initiation and thaw completion, eliminating subjectivity of determining the end point. ThawSTAR thawing instrument accepts vials from liquid nitrogen storage or from -80°C storage and delivers highly reproducible thermal profiles that mirror those obtained from a “perfect” water bath thaw, without the variability or risk of contamination.

ThawSTAR™ Cell Transport and Thawing System

ThawSTAR™ Transport and Thawing System vs. Water Bath: Performance Data

Ex-vivo stimulated B cells: A total of 6 vials, 2 each from 3 donors, were frozen in a -80°C freezer using a CoolCell® module and then stored in liquid nitrogen for two weeks. One vial from each donor was removed from LN2, placed into a ThawSTAR™ CFT Transporter and thawed in a ThawSTAR™ automated cell thawing instrument. The other vial from each donor was removed from LN2 and hand–carried to a water bath. The vials transported and thawed in the ThawSTAR transport and thawing system performed better in terms of cell viability and recovery. - Data generated by UCSF Diabetes Center and Transplant Surgery

To view a presentation by UCSF, visit: www.biocision.com/thawstar-video

Problem: Water Baths and Other Manual Thawing Methods• User dependent• Contamination risk• Variable end point• Not standardized

Solution: ThawSTAR™ Automated Cell Transport and Thawing System • Highly reproducible• Hands-free operation• No contamination• Standardized method

ThawSTAR™ Automated Cell Thawing System: Performance Data

How It Works

1.8 mL cryogenic vials filled with 1.0 mL of cryopreservation medium (10% DMSO/20% FBS/70% DMEM) were fitted with an interior wall thermocouple (< 0.5 mm from internal wall surface) and a central axis thermocouple, frozen at -80°C, and maintained at -75°C in a CoolRack® on dry ice. Three vials were thawed in a 37°C water bath (left panel) or in the ThawSTAR™ System (right panel). The temperature profiles recorded were very similar for both the water bath and ThawSTAR thaw. For the water bath thaw, the vials were removed from the bath when a pea-sized ice chunk remained (arrow) and then gently tapped to melt the chunk. Similarly, ThawSTAR system ejected the vial at the point where a pea-sized ice chunk remained (arrow) and after mixing the final vial temperature is ~5-10°C. Note that the sharp rise in temperature seen with the central thermocouple is indicative of the ice chunk breaking away from the thermocouple.

37°C Water Bath ThawSTAR

ThawSTAR™ Automated Cell Thawing System is an intuitive, error-free method for achieving reproducible thawing and recovery results. Simply insert a frozen cryogenic vial and retrieve when the vial is gently raised at the end of the thaw cycle. The automatic release of the vial, coupled with audio and visual alarms, allow quick removal of the thawed vial for downstream processing and minimize the risk of toxicity from cryopreservatives, such as DMSO. ThawSTAR thawing system delivers results comparable to those achieved when thawing a frozen vial in a 37°C water bath, but with reproducibility and standardization built in. Its small footprint enables easy incorporation into a laboratory hood or clinical setting.

ThawSTAR™ CFT Transporter is comprised of an outer insulative foam lid and base, and an internal round CoolRack® CFT5 sample module that accommodates up to five 1.8 or 2.0 mL cryogenic vials. Prior to using the ThawSTAR CFT Transporter, a thin layer of dry ice is added to the bottom of the base. The round CoolRack CFT5 module is placed over the dry ice and allowed to equilibrate to dry-ice temperature (approximately 10 minutes). Frozen vial is then extracted from storage and quickly placed into the module. The lid is placed securely on top of the base and all samples are maintained at dry ice temperature until transported to the thawing location.

Lid

CoolRack® CFT5

Base

Handle

Mean viability is compared for paired peripheral blood mononuclear cells (PBMCs) isolated immediately after phlebotomy using traditional Ficoll and SepMate methods (left). Mean viability is compared post cryopreservation of paired PBMCs isolated immediately after phlebotomy for cells thawed in a water bath and using ThawSTAR™ system (right). With fresh isolates, duplicate samples were processed by two different technicians to compare intertechnician variability for both PBMC isolation methods (results not shown) (n = 6). For cryopreserved samples, one vial from each of three donors was thawed for each condition being analyzed (n = 3).

- Data generated by Blood Systems Research Institute

Mean recovery is compared for paired PBMCs isolated immediately after phlebotomy using traditional Ficoll and SepMate methods (left) (n = 6). Mean recovery post cryopreservation is compared for paired PBMCs isolated immediately after phlebotomy for cells thawed in a water bath and using the ThawSTAR™ System (right) (n = 3).

- Data generated by Blood Systems Research Institute

Fresh CryopreservedFresh Cryopreserved

Thermal Profile Compared to Water Bath

Cell Viability Compared to Water Bath Cell Recovery Compared to Water Bath

Water Bath ThawSTAR™ Water Bath ThawSTAR™

Thawing Vials Directly from LN2 Stores

1.8 mL cryogenic vials filled with 1.0 mL of cryopreservation medium (10% DMSO/20% FBS/70% DMEM) were fitted with an interior wall thermocouple (< 0.5 mm from internal wall surface) and a central axis thermocouple and frozen at -196°C. The vials were removed from LN2 and immediate placed in the ThawSTAR™ instrument. Upon thaw completion, the vial was ejected at the point where a pea-sized ice chunk remained (arrow). Note that the sharp rise in temperature seen with the axial thermocouple is indicative of the ice chunk breaking away from the thermocouple. Final vial temperature is ~5-10°C, comparable to a water bath thaw.

Thawing Compared to Dry Bead Bath or Heat Block

1.8 mL cryogenic vials filled with 1.0 mL of cryopreservation medium (10% DMSO/20% FBS/70% DMEM) were fitted with a wall thermocouple (< 1 mm from internal wall surface) and a central axis thermocouple, frozen at -80°C, and maintained at -75°C in a CoolRack® on dry ice. Vials were thawed in either a ThawSTAR™ System (green traces), a 37°C bead bath (Lab Armor beads; red traces), or an aluminum heat block equilibrated to 37°C (blue traces). The ThawSTAR System thaw time is 2-3X faster than these other dry thawing methods.

ThawSTAR™ Automated Cell Thawing System: Reproducibility

Six vials were thawed in a 37°C water bath. The average thaw time in the water bath was 151 s, with a 99% confidence interval of 139-164 s (range shown as dotted lines). Left panel: The average time for the same ThawSTAR™ System to thaw > 5 frozen vials was measured each day for 5 days. Right panel: The average thaw time for three different ThawSTAR Systems was measured using > 5 frozen vials per unit on the same day. No significant differences were identified for either scenario at p<0.05 (2-way ANOVA with post hoc Sidak test).

3 ThawSTAR units tested on same day

99% conf. interval of water bath thaw time

Same ThawSTAR unit tested on 5 days

ThawSTAR™ CFT Transporter: Performance Data

ThawSTAR™ CFT Transporter holds cryogenic vials at near dry ice temperatures prior to thawing in the ThawSTAR™ System. Dry ice is placed in the foam holder below the metal CoolRack® module. To verify the holding temperature, a cryogenic vial filled with cryopreservative solution with a centrally-located thermocouple was frozen in LN2 then transferred to the pre-equilibrated CFT Transporter. The temperature profile shows that the vial equilibrates quickly (< 10 min) to the holding temperature and remains stable for > 1 hr.

© 2015. BioCision, LLC. All rights reserved. Patents pending. BioCision, ThawSTAR, STAR, CoolCell, CoolRack designations are trademarks owned by BioCision, LLC. PN 20056 Rev 1BioCision, LLC 101 Glacier Point Road, Suite E, San Rafael, CA 94901 USA sales@biocision.com www.biocision.com

®

Ordering Information

Specifications

The ThawSTAR™ System is for laboratory research use only. Any intended use for diagnostic purposes, direct transfusion, or in the production of therapeutic product(s) or vaccines(s) may require advance regulatory clearance which is the sole responsibility of the user, as this is not a medical device that has undergone medical device registration, clearance, or approval by the U.S. Food and Drug Administration (FDA), European Union, Health Canada, or the Australian Therapeutic Goods Administration. Research Only Device: Limited by Federal Law (United States) to Research Use Only.

ThawSTAR™ Transport and Thawing System A complete solution for standardized transport and thawing of frozen cells. Components include ThawSTAR Automated Cell Thawing System and ThawSTAR CFT Transporter.

Item No. DescriptionBCS-603 ThawSTAR Confirmation Vials

ThawSTAR™ Confirmation VialsFor use with ThawSTAR Automated Cell Thawing System before and after each thaw session to document instrument performance and audit trail. Recommended for single use only to minimize risk of cross-contamination. Pack of 20 vials.

For extended warranty information please contact us at +1.415.785.8516 or at support@biocision.com

Item No. DescriptionBCS-600 ThawSTAR Transport and Thawing System, complete

Includes: ThawSTAR CFT Transporter and ThawSTAR Automated Cell Thawing SystemBCS-601 ThawSTAR Automated Cell Thawing SystemBCS-602 ThawSTAR CFT Transporter

ThawSTAR™ Automated Cell Thawing SystemDimensions (Diameter x Height) 11.0 cm x 14.5 cm (5.7 in x 4.3 in)Thawing temperature Equivalent to a 37°C water bathThawing time Typically less than 3 minutesOperating environment temperature 5°C to 35°C; 75% room humidity (maximum)Storage temperature -20°C to 60°CMaximum altitude 4,000 mPower rating 40 W maximumInput/Output voltage (power adaptor) AC 100 - 240V / DC 12 V, 50 – 60 HzOutput current 5AElectrical plug formats US, UK, EUWeight without power supply 0.7 kg (1.6 lbs)Weight with power supply 1.2 kg (2.6 lbs)Compliance CE mark (EMC and Safety), IEC60601, IEC61010, RoHS, WEEE

Cryogenic Vial CompatibilityVial body external diameter 12.19 - 12.37 mm (0.480 - 0.487 in) *Measured 30.4 mm (1.2 in) from bottom of vialVial body external height below cap ring (minimum) 26.6 mm (1.05 in)Overall height of vial with cap (minimum) 44 mm (1.73 in)Vial cap diameter (maximum) 14.3 mm (0.56 in)Vial capacity One standard 1.8 to 2.0 mL cryogenic vialSample volume Optimized for 1.0 mL fill volumeVial contents Optimized for cells in cryopreservation medium such as 10% DMSO / 20% FBS / 70% DMEMVial label thickness allowed (maximum, if using) 0.1 mm

ThawSTAR™ CFT Transporter Dimensions (Diameter x Height) 11.4 x 15.0 cm (4.5 x 6.0 in)Composition Highly insulative cross-linked, closed cell polyethylene foamDry ice used ~150 gHolding temperature < -70°CHolding time (lid on) >1 hour *vial frozen in LN2 and placed in a pre-equilibrated CoolRack module and placed over ~150 g dry

ice in a CFT TransporterVial capacity Up to five 1.8 to 2.0 mL cryogenic vials