CoolCell Performance Data

CoolCell controlled-rate cell freezing containers are suitable for a variety of cell types and are currently being used in research, cell banking and clinical trials.

 

Cell types include: stem cells, primary cells, PBMCs, cell lines, insect cells, yeast, bacteria
Vessels include: 2ml through 5ml cryogenic vials, 15mm through 23mm diameter injectable ampules

CoolCell containers are cited in a number of protocols and journals.

Protocols
Publications

 

CoolCell® Freezing Profile Consistency

Cell freezing performance & consistency

Performance test: A temperature probe was placed into a 2.0 mL cryogenic vial containing 1.0 mL of water and the tube was inserted into a CoolCell® sitting at room temperature. The CoolCell was then placed directly into a -80°C freezer and the temperature rate and profile was observed over a 3 hour period. This experiment was repeated 5 consecutive times and temperature profiles were recorded.

Conclusion: BioCision’s CoolCell controlled-rate cell freezing module showed identical fusion time and cooling profiles over five consecutive freeze cycles.

CoolCell® vs. Alcohol-filled Container Freezing Profile

Alcohol-free cell freezing performance graph

Performance test: In an attempt to characterize the time lag and temperature rise of samples undergoing the freezing step, the following experiment was performed using a standard alcohol-filled freezing container and an insulated low-thermal mass cell freezing container (CoolCell). These two containers were continuously monitored for sample temperature in both room temperature and pre-chilled starting conditions.

Conclusion: In both starting conditions the alcohol-filled system warmed the pre-chilled samples more and longer than CoolCell before freezing overtakes initial sample temperature. The insulated CoolCell cryopreservation module had significantly lower heat stress on the samples and this will improve sample viability.

Download the full experiment: CoolCell vs. Isopropanol-filled container

Download: The CoolCell Advantage

CoolCell® FTS30 Freezing Profile

CCFTS30 cell freezing profile

Performance test: A thermocouple temperature probe was placed into 3 separate 2.0 mL cryogenic vials containing 1.0 mL of freezing media. Beginning at room temperature, the tubes were placed into a CoolCell FTS30 at various locations in the FTS module. The CoolCell FTS30 was then placed directly into a -75°C freezer and the temperature of the freezing media was recorded at 10 second intervals over a 4 hour period. The experiment was repeated 3 consecutive times

Conclusion: BioCision’s CoolCell FTS30 controlled-rate cell freezing module provides consistent vial to vial freezing and fusion duration cycle after cycle. In addition, the freezing profile is identical to that obtained with the widely accepted CoolCell 12 which has become the gold standard for cell freezing in thousands of laboratories.

Download the full experiment: CoolCell FTS30 freezing profile.

CoolCell® 5ml Freezing Profile with Varying Volumes of Media

CoolCell 5ml cryopreservation profile

Performance test: Beginning at room temperature, 5.0 mL cryogenic vials containing varying amounts of freezing media from 3.0 mL - 5.0 mL were placed into a CoolCell 5ml and this unit was then placed directly into a -75°C freezer. Using a thermocouple temperature probe the temperature of the freezing media was recorded at 10 second intervals over a 3.5 hour period.

Conclusion: BioCision’s CoolCell 5ml rate controlled freezing module provides consistent vial to vial freezing and fusion duration cycle after cycle, regardless of the volume of media in the 5.0 mL cryogenic vial - consistently showing highly reproducible controlled freezing rate of -1°C/min through until fusion.

Download the full experiment: CoolCell 5ml freezing profile

 

CoolCell® Performance: Stem Cells

Performance test: Effects of freezing on antigen-specific Treg (Ag-Treg) cell therapy products; Ag-Tregs (n = 6) were frozen at a concentration of 1 to 10 × 106 cells/mL using the CoolCell freezing device or programmable freezer (freezing rate of –1 °C/min). Viability and absolute viable cell count of thawed Ag-Treg cell therapy products were evaluated by flow cytometry.

See the poster presented at International Society for Cell Therapy >

 

Post-thaw viability of Human Embryonic Stem Cells (RC-10): 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 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. Data provided by Roslin Cellab, Scotland.

 

Cell Proliferation of Human Embryonic Stem Cells (RC-10): 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.) Data provided by Roslin Cellab, Scotland.

 

Differentiation of Human Embryonic Stem Cells 1 Day Post-Thaw: 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).  Data provided by Roslin Cellab, Scotland.

 

CoolCell Performance: Primary Cells and PBMCs

Recovery of PBMCs after cryopreservation: Average % of cell recovery after freezing with the CoolCell method was 77% compared to 72 % with the programmable freezer. Thus there was no significant difference between the 2 freezing methods. Data performed by UCSF Diabetes Center.

Conclusion: Comparison of freezing methods. Graph comparing % of cell recovery after freezing with the CoolCell freezing device (blue) versus freezing using a programmable step-down freezer (red) in 3 different samples at high cell concentration.

See the poster presented at International Society for Cell Therapy >

 

Viability of PBMCs after cryopreservation: Average % of cell viability after freezing with the CoolCell method was 97.6% compared to 98.6% with the programmable freezer. Thus there was no significant difference between the 2 freezing methods. Data performed by UCSF Diabetes Center.

Conclusion: Comparison of freezing methods. Graph comparing % of cell viability after freezing with the CoolCell freezing device (blue) versus freezing using a programmable step-down freezer (red) in 3 different samples.


See the poster presented at International Society for Cell Therapy >

 

Post-thaw viability of Peripheral Blood Mononuclear Cells (PBMC): PBMC cells were isolated and suspended in human serum with 10% DMSO to a concentration of 1x107 cells. Two cryogenic vials of each were prepared. One vial was frozen overnight using a BioCision CoolCell and one in a commercial IPA-based freezing system (“Mr. Frosty” by Thermo Fisher Scientific, Inc.). Following long term storage in LN2, cells were thawed and viability counts for each vial were obtained by the trypan blue stain method. Data performed by Stanford University.

 

Post-thaw viability of Human Umbilical Vein Endothelial Cells (HUVEC): One mL aliquots of cells resuspended in freezing medium at a concentration of 2x106 cells/ml. were proportioned into 1.8 mL cryogenic vials and frozen in either a BioCision CoolCell or in an alcohol-filled freezing unit (“Mr. Frosty” by Thermo Fisher Scientific, Inc.). Five vials frozen by either method were rapidly thawed and resuspended in growth media. Live cell counts were obtained by the trypan blue exclusion method. Data provided by Stanford University.

 

Post-thaw viability of PBMCs; Comparison of CoolCell, Electric Controlled-Rate Freezer & “Mr. Frosty”: Approximately 1.5 x 106 PBMCs suspended in cryopreservative were aliquoted into cryogenic vials and frozen in a BioCision CoolCell, an isopropanol-based container (“Mr. Frosty” by Thermo Fisher Scientific, Inc.) and in a programmable controlled-rate freezer.  The cells were thawed and counted using the Trypan Blue method. Each method yielded greater than 94% viability with no statistical difference between any of the methods. Data provided by ZenBio.

 

CoolCell Performace: Immortalized Cell Lines

Four cell lines – HeLa, CHO-K, K562 and NIH3T3 – were frozen in isopropanol-based “Mr. Frosty” freezing container, CoolCell 12-well container and CoolCell FTS30 30-well container. Alcohol-free CoolCell performed as well or better for each cell line in viability, GFP transfection efficiency and growth performance 24 hours post-thaw.  Data performed by Lonza, Germany.  “Mr. Frosty” is a Trademark of Thermo Fisher Scientific, Inc