G Rex Cell Culture

Conclusions We have successfully developed a static culture protocol for largescale expansion of NK cells in the gas permeable GRex system under good manufacturing practice (GMP) conditions This strategy is currently being used to produce NK cells for cancer immunotherapy.

G rex cell culture. Watch researchers from Baylor College of Medicine's Cell and Gene Therapy Center harvest 3 billion CART cells in only 100mls of culture medium The entire 1. ( http//wwwabnovacom ) Cell culture is the process by which cells are grown under controlled conditions This video shows you how to culture suspension. This cell density is much lower than the density range of 125–500 × 10 3 cells per cm 2 used in G‐Rex cultures of K562 cells, T‐cells, and other cell types 30, 35 G40D cultures generated 39 ± 19 CD41 CD42b Mks per input CD34 cell, which was higher than our current process with (C11D) or without (C11) a fed‐batch strategy at 28 ± 9 and 18 ± 8, respectively.

The use of GRex M devices in combination with our newly optimized protocol allows the operator to initiate a suspension culture with a small number of cells (125 × 10 5 cells/cm 2 ) and achieve a 100fold expansion in only 10 days using just 10 ml of culture medium per cm 2 significantly exceeding that previously achieved in tissue culture plates or bags 13,,21,25 Furthermore, since cells were supplemented with medium as a onetime upfront addition, we were able to utilize glucose. Wilson Wolf invents, develops, and manufactures superior devices for use in T Cell Therapy A current focus is on the evolution of our GRex® platform to create the best scale up and scale out. Using the elegant GRex culture platform, cellular therapies can be scaled up to bring immunotherapy from the benchtop to the bedside and treat large patient.

For scalingup adherent cells, Corning® CellSTACK®, provides a straightforward scaleup path However, some of our customers can’t accommodate larger stacks. Key to the GRex gas permeable cell culture process is the Liquid Silicone Rubber (LSR) 010 thick membrane molded by Minnesota Rubber and Plastics In the device, the LSR membrane provides convection nutrient transport (not diffusion) generating the most cell cultures in the least amount of space Convection is the key to achieving maximum cell growth with uninterrupted access to nutrients. The GRex is the only cell culture system capable of supporting significant cell expansion (100fold) with a single upfront addition of medium We demonstrated this to be true not only for K562 cells but also for primary T cells genetically modified to express a chimeric antigen receptor (Supplementary Figure S1) This unique characteristic allows not only a costeffective use of resources but also the measurement of elements within the media that predict the kinetics of cell growth.

Https//wwwthermofishercom/global/en/home/references/gibcocellculturebasicshtml?cid=BID_R01_PJT3313_BID8_VI_YUT_OD_KT_365The handbook and videos pr. GRex® 500MCS Cell Culture Bottle (CARCZ50) The surface area of 500 square centimeters permeable membrane has a medium capacity of 5000 milliliters Closed system for semiautomatic liquid handling Without changing the medium, 250 million cells can be expanded to 10 to billion cells in about 10 days GAMMA radiation. Culture supernatant was assessed for metabolite levels The remaining seeded cells were harvested and distributed into GRex culture devices at the equivalent seeding density of 1 × 10 6 cells/cm 2 in GRex 500MCS (Wilson Wolf, St Paul, MN, USA) On day 12, the GRex 500MCS was harvested and analyzed for cell count, viability, and phenotype.

Specification Name Specification Value;. The GRex cell culture platform is based on a gaspermeable membrane technology that provides numerous advantages over other systems Conventional bioreactor platform technologies developed for large scale mammalian cell expansion are typically constrained by the mechanics of delivering oxygen to an expanding cell population. Specification Name Specification Value;.

( http//wwwabnovacom ) Cell culture is the process by which cells are grown under controlled conditions This video shows you how to culture suspension. Wilson Wolf’s GRex ® cell culture technology, Fresenius Kabi’s Lovo and upcoming Cue cell processing systems, and BioTechne’s range of GMP proteins, reagents, media, and gene editing. EBVCTL harvested from GRex Collect 1000E06 Cell Approx 4 minutes process Minimal manipulation GRex decrease production cost 24 well plate24 well plate GRex Starting cell # 2x10 7 PBMCs 2x107 PBMCs Final cell # 1x1010 CTLs 1x1010 CTLs Tech time 129 hours $ 3,096 25 hours $ 60 GMP charge $ 2280 $ 300 Cultureware 310 plates $ 2325 2 (GRex40).

A singleuse bioreactor with novel design and features to accommodate modern cell culture processes Duration 607 Thermo Fisher Scientific 1,496 views 607 T cell expansion GRex vs rocking. Package Contents GRex 6 Well Culture Plate, 35ml Number of Wells 6 Sterility Gamma Irradiated Material Gas Permeable Membrane. Package Contents GRex 6 Well Culture Plate, 35ml Number of Wells 6 Sterility Gamma Irradiated Material Gas Permeable Membrane.

Using the GRex culture flask supplied by Wilson Wolf, we can now readily grow sufficient quantities of these cells for therapeutic purposes The GatheRex pump which we are requesting is the only system compatible with the Grex culture flasks and allows for the concentration and harvesting of the Tcells at the end of the culture. GRex® MultiWell Cell Culture Plates, Argos Technologies GRex® multiwell technology is ideal for applications involving nonadherent cells Compared to traditional multiwell plates, advantages of the GRex® multiwell plates include nutrient availability that is uncoupled from oxygen delivery leading up to 100fold cell expansion in less time with fewer manipulations. The Xuri Cell Expansion System W25 delivers an automated, controlled expansion environment for the growth of cell therapies in volumes up to 25 L across research, process development, and manufacturing operations Closed system minimizes risk of cell contamination High cell densities through automated perfusion protocols.

The GRex cell culture platform is based on a gaspermeable membrane technology that provides numerous advantages over other systems Conventional bioreactor platform technologies developed for large scale mammalian cell expansion are typically constrained by the mechanics of delivering oxygen to an expanding cell population. Cells were cultured in TexMACS GMP medium with interleukin (IL)7 and IL15 and transduced with RetroNectin in bags or Vectorfusin1 in the GRex Total viable cell number, fold expansion, viability, transduction efficiency, phenotype and function were compared between the two processes. Key to the GRex gaspermeable cell culture process is a thin liquid silicone rubber (LSR) membrane molded by Minnesota Rubber and Plastics In the device, the LSR membrane provides convection nutrient transport (not diffusion), generating the most cell cultures in the least amount of space.

The GRex cell culture vessels allow gas exchange across the base of the culture, thus supporting higher cell number per unit surface area and increased cell survival and expansion rates The GRex vessels have been tested for the expansion of several types of immune cells. Collection of Cells from Culture Collect medium from the culture and place in a tube Immediately add PBS to the adherent cells Pipet up and down to rinse the culture surface and pool this with the cell in medium Add PBS to the culture and place the culture in the incubator at 37°C for 10–15 minutes During this time, the cells should begin to detach from the culture surface. Using the GRex culture flask supplied by Wilson Wolf, we can now readily grow sufficient quantities of these cells for therapeutic purposes The GatheRex pump which we are requesting is the only system compatible with the Grex culture flasks and allows for the concentration and harvesting of the Tcells at the end of the culture.

In order to harvest the cells expanded in the closed system GRex without exposing the culture to the environment, Wilson Wolf developed the “GatheRex,” a semiautomated system that allows the operator to drain the excess media present in the culture and collect cells without risk of contamination”. GRex® stands for Gas Permeable Rapid Expansion and is the result of applying our disruptive technology to the needs of immunotherapy GRex® is the only production platform created specifically to produce immune cells such as T cells, natural killer cells, and hematopoietic cells GRex® is proving to be the most practical and cost effective way to scale up and scale out. Key to the GRex gas permeable cell culture process is the gas permeable membrane molded by Minnesota Rubber and Plastics In the device, the gas permeable membrane provides convection nutrient transport (not diffusion) generating the most cell cultures in the least amount of space.

We optimized the coculture ratio of K562 aAPCs to immune cells, and migrated this method to a GRex cell growth platform to derive clinically relevant cell numbers in a Good Manufacturing Practice (GMP)compliant manner We further include a depletion step to selectively remove αβ T lymphocytes. The GRex cell culture platform is based on a gaspermeable membrane technology that provides numerous advantages over other systems Conventional bioreactor platform technologies developed for. Collection of Cells from Culture Collect medium from the culture and place in a tube Immediately add PBS to the adherent cells Pipet up and down to rinse the culture surface and pool this with the cell in medium Add PBS to the culture and place the culture in the incubator at 37°C for 10–15 minutes During this time, the cells should begin to detach from the culture surface.

Using the GRex culture flask supplied by Wilson Wolf, we can now readily grow sufficient quantities of these cells for therapeutic purposes The GatheRex pump which we are requesting is the only system compatible with the Grex culture flasks and allows for the concentration and harvesting of the Tcells at the end of the culture. EBVCTL harvested from GRex Collect 1000E06 Cell Approx 4 minutes process Minimal manipulation GRex decrease production cost 24 well plate24 well plate GRex Starting cell # 2x10 7 PBMCs 2x107 PBMCs Final cell # 1x1010 CTLs 1x1010 CTLs Tech time 129 hours $ 3,096 25 hours $ 60 GMP charge $ 2280 $ 300 Cultureware 310 plates $ 2325 2 (GRex40). The GRex devices should be at maximum capacity by now But if they are slow growing, you may have to feed again as previously described for the steps on Day 15 and continue the culture Repeat medium exchanges and cytokine addition as needed until the cells have reached a maximum number at high viability Collect the cells from the GRex.

GRex® 100MCS Cell Culture Bottle GRex® 100MCS Cell Culture Bottle (CARCZ54) The surface area of 100 square centimeters of breathable membrane, the filling volume is 1000 ml Closed system for semiautomatic liquid handling In about 10 days, 50 million cells can be expanded to between 2 and 4 billion cells without medium exchange. The GRex 100 is a production platform designed specifically to produce immune cells With our experience in the medical device field, we evaluated the product design from a fresh perspective Along with completing the design for the compression seal, we enhanced the sealing of the packaging. The GRex cell culture vessels allow gas exchange across the base of the culture, thus supporting higher cell number per unit surface area and increased cell survival and expansion rates The GRex vessels have been tested for the expansion of several types of immune cells.

The GRex is the only cell culture system capable of supporting significant cell expansion (100fold) with a single upfront addition of medium We demonstrated this to be true not only for K562 cells but also for primary T cells genetically modified to express a chimeric antigen receptor (Supplementary Figure S1) This unique characteristic allows not only a costeffective use of resources but also the measurement of elements within the media that predict the kinetics of cell growth. Culture supernatant was assessed for metabolite levels The remaining seeded cells were harvested and distributed into GRex culture devices at the equivalent seeding density of 1 × 10 6 cells/cm 2 in GRex 500MCS (Wilson Wolf, St Paul, MN, USA) On day 12, the GRex 500MCS was harvested and analyzed for cell count, viability, and phenotype. The GRex cell culture platform is based on a gaspermeable membrane technology that provides numerous advantages over other systems Conventional bioreactor platform technologies developed for large scale mammalian cell expansion are typically constrained by the mechanics of delivering oxygen to an expanding cell population These systems often utilize complex mechanisms to enhance oxygen delivery, such as stirring, rocking, or perfusion, which adds to expense and increases their overall.