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CoolCell® DL Cell Freezing Containers: Controlled-Rate Passive Freezing Containers with Data Logging for Validatable Processing
S84
Poster Abstracts
cryogenic vial format. This vial format is commonly used in both research and clinical settings, making the ThawSTAR CFT2 a useful tool for de-risking thawing of clinical or translation research samples. Building on broad interest in the ThawSTAR Platform, BioCision has introduced additional formats (the AT6 for 6 ml cell therapy vials and the NG1.5 for 1.5 ml cryovials) and improved the ThawSTAR Platform functionality with next generation features (improved load detection, RFID reading, connectivity). The ThawSTAR AT6 utilizes the same proprietary thermal sensing technology of our first generation CFT2 system, and extends that technology to a system capable of thawing between 2 and 6 ml. In addition, this second generation format incorporates upgraded hardware and software that results in better load detection and thaw time determination. Celgene plans to use this format for their Phase 2 clinical trial work on an allogeneic cell therapy to treat symptoms of diabetes. The ThawSTARTM Automated Cell Thawing Platform provides users within research and clinical settings a thawing solution that is intuitive, standardized, reproducible and traceable.
Figure: BioCision’s CoolCell DL combines highly reproducible controlled rate freezing with data logging capabilities. Image Credit: BioCision 242 DL CELL FREEZING CONTAINERS: COOLCELL ® CONTROLLED-RATE PASSIVE FREEZING CONTAINERS WITH DATA LOGGING FOR VALIDATABLE PROCESSING E.J. Kunkel, B. Schryver, M. Thompson, R. Ehrhardt BioCision LLC, San Rafael, California, United States The potential of cell-based research for drug discovery and therapeutic treatment is well recognized. Successful translation of therapeutic cell processing from a research lab to a clinic, however, requires handling procedures and preservation technology that guarantee quality, reproducibility, and safety of the final product. To ensure consistent, standardized freezing of therapeutic cells for long term storage, many labs make use of a dedicated, controlled rate freezer to minimize variability during the freezing process. Though they are certainly effective and have built-in validation procedures, controlled rate freezers (CRFs) do have significant drawbacks: they are expensive, have a large footprint, and can be difficult to maintain and operate. Passive freezing devices provide a low-cost, scalable alternative to controlled rate freezers, but in certain situations cannot be taken advantage of due to regulatory specifications regarding tracking and archiving of temperature data. This problem has now been addressed by BioCision’s CoolCell® DL (Data Logger) cell freezing container. BioCision’s line of CoolCell freezing containers provide highly reproducible, alcohol-free 1°C/minute controlled rate freezing when placed in a −80°C freezer. They are proven for use with stem cells, primary cells, and cell lines, and have already been adopted for use in GMP regulated workflows. The CoolCell DL container will unite these features with NIST traceable temperature logging and data exporting capabilities. Temperature measurements collected at regular intervals during the cell freezing process can be recorded, then easily downloaded to a personal computer via a supplied USB cable. CoolCell DL provides an economic alternative to large, expensive CRFs, while the unit’s data logging capabilities ensure that any cryopreservation steps during the translational process are consistent and standardized across multiple pre-clinical and clinical sites.
243 FACTORS AFFECTING CLINIMACS ENRICHMENT OF CD34+ CELLS FROM PERIPHERAL BLOOD STEM CELL PRODUCTS: A SINGLE CENTER ANALYSIS OF 278 PROCEDURES P. Law2, B. Triplett1, W. Leung1, G. Kang3, W. Janssen2,1 1 Bone Marrow Transplantation & Cellular Therapy, St Jude Children’s Research Hospital, Memphis, Tennessee, United States, 2Human Applications Laboratory, St Jude Children’s Research Hospital, Memphis, Tennessee, United States, 3Biostatistics, St Jude Children’s Research Hospital, Memphis, Tennessee, United States Enrichment of CD34+ cells using CliniMACS has received FDA approval for selected clinical indications. The process has been in use (clinical trials plus routine applications) for over 15 years at St Jude Children’s Research Hospital. In this abstract we report our single institution analysis of 278 CD34+ cell enrichment procedures since 2000 using mobilized peripheral blood stem cell (PBSC) products. The manufacturer’s procedures were followed and the final products were either infused directly or cryopreserved according to various clinical trials. A summary of the results is shown in the Table. Correlation analyses show that purity (%CD34+ cells) of final product is directly correlated to % CD34+ cells in the apheresis unit, as expected and confirming earlier reports. Furthermore, when the data are analyzed using a general multivariate regression model, high purity can be predicted by (in order of decreasing importance): low platelets (plt), low hematocrit HCT), high mononuclear cells (MNC), and low total nucleated cells (TNC). High recovery of CD34+ cells, on the other hand, could be predicted by (in order of deceasing importance): high MNC, low HCT, low plt, and low TNC. In conclusion, our data showed that the apheresis should be carefully performed to ensure the PBSC collection contained high proportion of MNC with as few plt & RBC as possible. Also, all cell washes (prior to and after bead labeling) should be performed at low centrifuge speed to remove as much plt as possible. If the apheresis unit has high plt content, additional washes should be considered so that performance of CliniMACS enrichment is not compromised.
Table. Parameters affecting enrichment of CD34+ cells (median with ranges) Incoming Apheresis unit
All (278) Final Purity > 80% (249) Final Purity < 80% (29) CD34+ Rec > 70% (183) CD34+ Rec < 70% (95)
Final Product
TNC
% CD34+
% MNC
% HCT
plt after washes
% CD34+
Rec CD34+ cells
3.0 × 10^10 (0.9–15.8) 3.1 × 10^10 (1.1–15.8) 2.7 × 10^10 (0.9–6.7) 2.8 × 10^10 (0.9–7.3) 3.7 × 10^10 (0.9–15.8)
0.90% (0.1–4.46%) 0.93% (0.22–4.5) 0.57% (0.10–1.11) 0.89% (0.10–3.69) 0.91% (0.22–4.46)
92% (39–100) 92% (39–100) 79% (42–100) 92% (42–100) 92% (39–100)
3.8% (1.5–12.8) 3.7% (1.5–12.1) 4.3% (2.0–12.8) 3.7% (1.5–12.8) 3.9% (2.0–11.0)
3.0 × 10^10 (0.7–18.6) 3.1 × 10^10 (0.7–18.6) 2.1 × 10^10 (0.7–11.1) 2.6 × 10^10 (0.7–18.6) 4.0 × 10^10 (1.1–16.3)
94.1% (36.7–99.7) 95.1% (80.3–99.7) 69.8% (36.7–77.9) 94.1% (36.7–99.7) 94.3% (54.0–99.2)
74.7% (23.6–125) 74.8% (23.6–125) 71.2% (51.7–100.8) 80.2% (70.1–125) 62.4% (23.6–69.5)
Poster Abstracts
cryogenic vial format. This vial format is commonly used in both research and clinical settings, making the ThawSTAR CFT2 a useful tool for de-risking thawing of clinical or translation research samples. Building on broad interest in the ThawSTAR Platform, BioCision has introduced additional formats (the AT6 for 6 ml cell therapy vials and the NG1.5 for 1.5 ml cryovials) and improved the ThawSTAR Platform functionality with next generation features (improved load detection, RFID reading, connectivity). The ThawSTAR AT6 utilizes the same proprietary thermal sensing technology of our first generation CFT2 system, and extends that technology to a system capable of thawing between 2 and 6 ml. In addition, this second generation format incorporates upgraded hardware and software that results in better load detection and thaw time determination. Celgene plans to use this format for their Phase 2 clinical trial work on an allogeneic cell therapy to treat symptoms of diabetes. The ThawSTARTM Automated Cell Thawing Platform provides users within research and clinical settings a thawing solution that is intuitive, standardized, reproducible and traceable.
Figure: BioCision’s CoolCell DL combines highly reproducible controlled rate freezing with data logging capabilities. Image Credit: BioCision 242 DL CELL FREEZING CONTAINERS: COOLCELL ® CONTROLLED-RATE PASSIVE FREEZING CONTAINERS WITH DATA LOGGING FOR VALIDATABLE PROCESSING E.J. Kunkel, B. Schryver, M. Thompson, R. Ehrhardt BioCision LLC, San Rafael, California, United States The potential of cell-based research for drug discovery and therapeutic treatment is well recognized. Successful translation of therapeutic cell processing from a research lab to a clinic, however, requires handling procedures and preservation technology that guarantee quality, reproducibility, and safety of the final product. To ensure consistent, standardized freezing of therapeutic cells for long term storage, many labs make use of a dedicated, controlled rate freezer to minimize variability during the freezing process. Though they are certainly effective and have built-in validation procedures, controlled rate freezers (CRFs) do have significant drawbacks: they are expensive, have a large footprint, and can be difficult to maintain and operate. Passive freezing devices provide a low-cost, scalable alternative to controlled rate freezers, but in certain situations cannot be taken advantage of due to regulatory specifications regarding tracking and archiving of temperature data. This problem has now been addressed by BioCision’s CoolCell® DL (Data Logger) cell freezing container. BioCision’s line of CoolCell freezing containers provide highly reproducible, alcohol-free 1°C/minute controlled rate freezing when placed in a −80°C freezer. They are proven for use with stem cells, primary cells, and cell lines, and have already been adopted for use in GMP regulated workflows. The CoolCell DL container will unite these features with NIST traceable temperature logging and data exporting capabilities. Temperature measurements collected at regular intervals during the cell freezing process can be recorded, then easily downloaded to a personal computer via a supplied USB cable. CoolCell DL provides an economic alternative to large, expensive CRFs, while the unit’s data logging capabilities ensure that any cryopreservation steps during the translational process are consistent and standardized across multiple pre-clinical and clinical sites.
243 FACTORS AFFECTING CLINIMACS ENRICHMENT OF CD34+ CELLS FROM PERIPHERAL BLOOD STEM CELL PRODUCTS: A SINGLE CENTER ANALYSIS OF 278 PROCEDURES P. Law2, B. Triplett1, W. Leung1, G. Kang3, W. Janssen2,1 1 Bone Marrow Transplantation & Cellular Therapy, St Jude Children’s Research Hospital, Memphis, Tennessee, United States, 2Human Applications Laboratory, St Jude Children’s Research Hospital, Memphis, Tennessee, United States, 3Biostatistics, St Jude Children’s Research Hospital, Memphis, Tennessee, United States Enrichment of CD34+ cells using CliniMACS has received FDA approval for selected clinical indications. The process has been in use (clinical trials plus routine applications) for over 15 years at St Jude Children’s Research Hospital. In this abstract we report our single institution analysis of 278 CD34+ cell enrichment procedures since 2000 using mobilized peripheral blood stem cell (PBSC) products. The manufacturer’s procedures were followed and the final products were either infused directly or cryopreserved according to various clinical trials. A summary of the results is shown in the Table. Correlation analyses show that purity (%CD34+ cells) of final product is directly correlated to % CD34+ cells in the apheresis unit, as expected and confirming earlier reports. Furthermore, when the data are analyzed using a general multivariate regression model, high purity can be predicted by (in order of decreasing importance): low platelets (plt), low hematocrit HCT), high mononuclear cells (MNC), and low total nucleated cells (TNC). High recovery of CD34+ cells, on the other hand, could be predicted by (in order of deceasing importance): high MNC, low HCT, low plt, and low TNC. In conclusion, our data showed that the apheresis should be carefully performed to ensure the PBSC collection contained high proportion of MNC with as few plt & RBC as possible. Also, all cell washes (prior to and after bead labeling) should be performed at low centrifuge speed to remove as much plt as possible. If the apheresis unit has high plt content, additional washes should be considered so that performance of CliniMACS enrichment is not compromised.
Table. Parameters affecting enrichment of CD34+ cells (median with ranges) Incoming Apheresis unit
All (278) Final Purity > 80% (249) Final Purity < 80% (29) CD34+ Rec > 70% (183) CD34+ Rec < 70% (95)
Final Product
TNC
% CD34+
% MNC
% HCT
plt after washes
% CD34+
Rec CD34+ cells
3.0 × 10^10 (0.9–15.8) 3.1 × 10^10 (1.1–15.8) 2.7 × 10^10 (0.9–6.7) 2.8 × 10^10 (0.9–7.3) 3.7 × 10^10 (0.9–15.8)
0.90% (0.1–4.46%) 0.93% (0.22–4.5) 0.57% (0.10–1.11) 0.89% (0.10–3.69) 0.91% (0.22–4.46)
92% (39–100) 92% (39–100) 79% (42–100) 92% (42–100) 92% (39–100)
3.8% (1.5–12.8) 3.7% (1.5–12.1) 4.3% (2.0–12.8) 3.7% (1.5–12.8) 3.9% (2.0–11.0)
3.0 × 10^10 (0.7–18.6) 3.1 × 10^10 (0.7–18.6) 2.1 × 10^10 (0.7–11.1) 2.6 × 10^10 (0.7–18.6) 4.0 × 10^10 (1.1–16.3)
94.1% (36.7–99.7) 95.1% (80.3–99.7) 69.8% (36.7–77.9) 94.1% (36.7–99.7) 94.3% (54.0–99.2)
74.7% (23.6–125) 74.8% (23.6–125) 71.2% (51.7–100.8) 80.2% (70.1–125) 62.4% (23.6–69.5)