Limits...
Optimizing the production of suspension cells using the G-Rex "M" series.

Bajgain P, Mucharla R, Wilson J, Welch D, Anurathapan U, Liang B, Lu X, Ripple K, Centanni JM, Hall C, Hsu D, Couture LA, Gupta S, Gee AP, Heslop HE, Leen AM, Rooney CM, Vera JF - Mol Ther Methods Clin Dev (2014)

Bottom Line: Broader implementation of cell-based therapies has been hindered by the logistics associated with the expansion of clinically relevant cell numbers ex vivo.A multicenter study confirmed that this fully optimized cell culture system can reliably produce a 100-fold cell expansion in only 10 days using 1L of medium.The G-Rex M series is linearly scalable and adaptable as a closed system, allowing an easy translation of preclinical protocols into the good manufacturing practice.

View Article: PubMed Central - PubMed

Affiliation: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston Methodist Hospital, Texas Children's Hospital , Houston, Texas, USA.

ABSTRACT
Broader implementation of cell-based therapies has been hindered by the logistics associated with the expansion of clinically relevant cell numbers ex vivo. To overcome this limitation, Wilson Wolf Manufacturing developed the G-Rex, a cell culture flask with a gas-permeable membrane at the base that supports large media volumes without compromising gas exchange. Although this culture platform has recently gained traction with the scientific community due to its superior performance when compared with traditional culture systems, the limits of this technology have yet to be explored. In this study, we investigated multiple variables including optimal seeding density and media volume, as well as maximum cell output per unit of surface area. Additionally, we have identified a novel means of estimating culture growth kinetics. All of these parameters were subsequently integrated into a novel G-Rex "M" series, which can accommodate these optimal conditions. A multicenter study confirmed that this fully optimized cell culture system can reliably produce a 100-fold cell expansion in only 10 days using 1L of medium. The G-Rex M series is linearly scalable and adaptable as a closed system, allowing an easy translation of preclinical protocols into the good manufacturing practice.

No MeSH data available.


Related in: MedlinePlus

Automated collection of cells in a closed G-Rex system. Panel (a) shows the setup of “GatheRex” draining the excess medium to concentrate the cells: (1) pump, (2) 0.2-µm sterile filter, (3) medium collection button, (4) medium line clamp, (5,6) medium/supplements input ports, (7) medium sampling port, (8) medium collection port, (9) medium bag, (10) medium, and (11) medium line optical detector. Panel (b) shows the GatheRex harvesting the concentrated cells: (12) resuspended cells, (13) cell collection button, (14) cell line clamp, (15) cell collection port, (16) cell collection bag, (17) harvested cells, and (18) cell line optical detector. Panel (c) shows a comparison in cell recovery achieved when cells are collected manually versus by the GatheRex. To confirm the absence of cells in cell collection tubing, the medium collection bag or the G-Rex following cell collection with the GatheRex, each component was washed with media and residual cells were harvested and counted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4362380&req=5

fig7: Automated collection of cells in a closed G-Rex system. Panel (a) shows the setup of “GatheRex” draining the excess medium to concentrate the cells: (1) pump, (2) 0.2-µm sterile filter, (3) medium collection button, (4) medium line clamp, (5,6) medium/supplements input ports, (7) medium sampling port, (8) medium collection port, (9) medium bag, (10) medium, and (11) medium line optical detector. Panel (b) shows the GatheRex harvesting the concentrated cells: (12) resuspended cells, (13) cell collection button, (14) cell line clamp, (15) cell collection port, (16) cell collection bag, (17) harvested cells, and (18) cell line optical detector. Panel (c) shows a comparison in cell recovery achieved when cells are collected manually versus by the GatheRex. To confirm the absence of cells in cell collection tubing, the medium collection bag or the G-Rex following cell collection with the GatheRex, each component was washed with media and residual cells were harvested and counted.

Mentions: The GatheRex (Figure 7a,b) operates via an air pump which pressurizes the G-Rex device with sterile air, allowing 90% of the medium residing above the cells to be displaced into a medium collection bag. Once this process is complete, optical detector #1 senses the presence of air in the medium collection line, automatically stopping the pump.


Optimizing the production of suspension cells using the G-Rex "M" series.

Bajgain P, Mucharla R, Wilson J, Welch D, Anurathapan U, Liang B, Lu X, Ripple K, Centanni JM, Hall C, Hsu D, Couture LA, Gupta S, Gee AP, Heslop HE, Leen AM, Rooney CM, Vera JF - Mol Ther Methods Clin Dev (2014)

Automated collection of cells in a closed G-Rex system. Panel (a) shows the setup of “GatheRex” draining the excess medium to concentrate the cells: (1) pump, (2) 0.2-µm sterile filter, (3) medium collection button, (4) medium line clamp, (5,6) medium/supplements input ports, (7) medium sampling port, (8) medium collection port, (9) medium bag, (10) medium, and (11) medium line optical detector. Panel (b) shows the GatheRex harvesting the concentrated cells: (12) resuspended cells, (13) cell collection button, (14) cell line clamp, (15) cell collection port, (16) cell collection bag, (17) harvested cells, and (18) cell line optical detector. Panel (c) shows a comparison in cell recovery achieved when cells are collected manually versus by the GatheRex. To confirm the absence of cells in cell collection tubing, the medium collection bag or the G-Rex following cell collection with the GatheRex, each component was washed with media and residual cells were harvested and counted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362380&req=5

fig7: Automated collection of cells in a closed G-Rex system. Panel (a) shows the setup of “GatheRex” draining the excess medium to concentrate the cells: (1) pump, (2) 0.2-µm sterile filter, (3) medium collection button, (4) medium line clamp, (5,6) medium/supplements input ports, (7) medium sampling port, (8) medium collection port, (9) medium bag, (10) medium, and (11) medium line optical detector. Panel (b) shows the GatheRex harvesting the concentrated cells: (12) resuspended cells, (13) cell collection button, (14) cell line clamp, (15) cell collection port, (16) cell collection bag, (17) harvested cells, and (18) cell line optical detector. Panel (c) shows a comparison in cell recovery achieved when cells are collected manually versus by the GatheRex. To confirm the absence of cells in cell collection tubing, the medium collection bag or the G-Rex following cell collection with the GatheRex, each component was washed with media and residual cells were harvested and counted.
Mentions: The GatheRex (Figure 7a,b) operates via an air pump which pressurizes the G-Rex device with sterile air, allowing 90% of the medium residing above the cells to be displaced into a medium collection bag. Once this process is complete, optical detector #1 senses the presence of air in the medium collection line, automatically stopping the pump.

Bottom Line: Broader implementation of cell-based therapies has been hindered by the logistics associated with the expansion of clinically relevant cell numbers ex vivo.A multicenter study confirmed that this fully optimized cell culture system can reliably produce a 100-fold cell expansion in only 10 days using 1L of medium.The G-Rex M series is linearly scalable and adaptable as a closed system, allowing an easy translation of preclinical protocols into the good manufacturing practice.

View Article: PubMed Central - PubMed

Affiliation: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston Methodist Hospital, Texas Children's Hospital , Houston, Texas, USA.

ABSTRACT
Broader implementation of cell-based therapies has been hindered by the logistics associated with the expansion of clinically relevant cell numbers ex vivo. To overcome this limitation, Wilson Wolf Manufacturing developed the G-Rex, a cell culture flask with a gas-permeable membrane at the base that supports large media volumes without compromising gas exchange. Although this culture platform has recently gained traction with the scientific community due to its superior performance when compared with traditional culture systems, the limits of this technology have yet to be explored. In this study, we investigated multiple variables including optimal seeding density and media volume, as well as maximum cell output per unit of surface area. Additionally, we have identified a novel means of estimating culture growth kinetics. All of these parameters were subsequently integrated into a novel G-Rex "M" series, which can accommodate these optimal conditions. A multicenter study confirmed that this fully optimized cell culture system can reliably produce a 100-fold cell expansion in only 10 days using 1L of medium. The G-Rex M series is linearly scalable and adaptable as a closed system, allowing an easy translation of preclinical protocols into the good manufacturing practice.

No MeSH data available.


Related in: MedlinePlus