Development of a single‐pass ceramic matrix bioreactor for large‐scale mammalian cell culture

Applegate, Mark A.; Stephanopoulos, Gregory

doi:10.1002/bit.260400909

Cited by 10 publications

(9 citation statements)

References 15 publications

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“…The bioreactors were constructed using ceramic monolith structures obtained from Corning Inc. (Corning, NY). The monoliths, described in detail by Applegate (1991), were composed of cordierite and were arranged with open square channels (1.5 mm width, 31 channels/cm 2 ) separated by macroporous ceramic walls (340 µm wall thickness, 20-22 µm mean pore size, 50% porosity). Other physical properties of note are the total pore volume (0.54 cm 3 /g of ceramic) and the bulk density (1.07 g/cm 3 ) of the monoliths.…”

Section: Sample Preparation For Radioimmunoassay (Ria)mentioning

confidence: 99%

“…Other physical properties of note are the total pore volume (0.54 cm 3 /g of ceramic) and the bulk density (1.07 g/cm 3 ) of the monoliths. The material was pretreated for biocompatibility by boiling with 10% nitric acid and rinsing with deionized water and PBS as described by Applegate (1991). Preliminary experiments demonstrated that TC-3 cells grew in the pores and channels of this material and that they remained responsive to secretagogue stimulus.…”

Section: Sample Preparation For Radioimmunoassay (Ria)mentioning

confidence: 99%

“…The design and operation of a single-pass, ceramic-bed bioreactor has also been described in detail previously (Applegate, 1991). Briefly, a ceramic monolith is arranged around small-scale flow channels through which are threaded oxygen-permeable tubes.…”

Section: Introductionmentioning

confidence: 99%

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Cyclic Operation of Ceramic‐Matrix Animal Cell Bioreactors for Controlled Secretion of an Endocrine Hormone. A Comparison of Single‐Pass and Recycle Modes of Operation

Grampp

Applegate

Stephanopoulos

1996

Biotechnology Progress

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Controlled secretion processes for the production of secretory proteins in monolayer culture have been described previously (Grampp et al. Adv. Biochem. Eng./ Biotechnol. 1992, 46, 35-62), but little is known about the feasibility of scaling such processes into high-density bioreactors. Two immobilized-cell, ceramic-matrix bioreactor configurations were tested using the beta TC-3 cell model system which, in monolayer culture, can be manipulated to secrete murine insulin in a highly controlled manner. One reactor was configured with an external recirculation reservoir for oxygen transfer and was operated as a conventional immobilized bed/recycle reactor. The other reactor was configured as a single-pass perfusion system with oxygen supplied by diffusion from silicone tubing positioned proximal to the porous walls of the ceramic matrix. After inoculation with beta TC-3 cells, both systems were perfused with serum-supplemented medium to stimulate cell growth, and they ultimately attained high densities (approximately 5 x 10(8) cells/mL of pore volume). To initiate controlled secretion operations, the reactor cores were washed with a serum-free basal medium, then exposed to a serum-free discharging medium containing secretory stimulants. Following several hours of discharging, the reactors were washed again, then switched to a serum-containing medium designed to quench the regulated secretion process. For the single-pass reactor these cycling operations were simple to implement and were effective in promoting the cyclic discharge and recharge of murine insulin. Because of the ability to reduce the perfusion rate in the single-pass reactor independent of oxygen transfer, the discharged insulin was captured in a relatively small volume (2 reactor core hold-up volumes), yielding a mean product concentration 10-fold greater than in the steady-state perfusate. Cyclic operation of the recirculating reactor was more difficult due to the complexity of switching between recirculation reservoirs, and the introduction of air bubbles during such operations resulted in the loss of biomass from the reactor after one cycle. Even in the first discharging cycle, the insulin yield was much lower than in the perfusate from the single-pass reactor, despite the comparable metabolic rates. The single-pass reactor was cycled successfully through four discharging and recharging episodes and maintained its ability to discharge insulin, albeit at a slower rate after the first discharge. Overall, 50-60% of the insulin secreted during the 48 h cycles was recovered during the brief discharging episodes. When insulin secretion rates and discharging yields were normalized to metabolic activity, neither high-density reactor system performed as well as did identically treated control T-flask cultures. It is hypothesized that the productivity and responsiveness of the high-density, pore-immobilized beta TC-3 cells are lower than in monolayer culture.

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Section: Sample Preparation For Radioimmunoassay (Ria)mentioning

confidence: 99%

Section: Sample Preparation For Radioimmunoassay (Ria)mentioning

confidence: 99%

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Cyclic Operation of Ceramic‐Matrix Animal Cell Bioreactors for Controlled Secretion of an Endocrine Hormone. A Comparison of Single‐Pass and Recycle Modes of Operation

Grampp

Applegate

Stephanopoulos

1996

Biotechnology Progress

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“…Although several novel continuous bioreactors have been developed for the culture of suspended mammalian cells (1)(2)(3)(4)(5)(6)(7)(8)(9), conventional roller bottles still provide the simplest means for culturing suspended mammalian cells that are shearsensitive but still require some means of agitation for cell suspension and oxygen and nutrient dispersion. Although each novel bioreactor has its own advantages, such as improvements in cell density, viability percentages, monoclonal antibody secretion rates, etc., these systems are generally not as mechanically simple or as easily scaleable as are roller bottles, require special user training, and require capital investment.…”

Section: Introductionmentioning

confidence: 99%

A Self-Feeding Roller Bottle for Continuous Cell Culture

Berson

Friederichs

2008

Biotechnol. Prog.

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The concept of a self-feeding roller bottle that delivers a continuous supply of fresh media to cells in culture, which is mechanically simplistic and works with existing roller apparatuses, is presented here. A conventional roller bottle is partitioned into two chambers; one chamber contains the fresh culture media reservoir, and the other contains the cell culture chamber. A spiroid of tubing inside the fresh media reservoir acts as a pump when the bottle rotates on its horizontal axis, continuously delivering fresh media through an opening in the partition to the cell culture chamber. The modified bottle proved capable of maintaining steady-state cell densities of a hybridoma cell line over the 10-day period tested, although at lower densities than reached during batch operation due to the continuous volume dilution. Steady-state density proved to be controllable by adjusting the perfusion rate, which changes with the rotation rate of the bottle. Specific antibody production rate is as much as 3.7 times the rate in conventional roller bottles operating with intermittent batch feeding.

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“…Continuous supply of medium coupled with continuous or intermittent cell or product harvest should result in a longer production phase and more efficient use of cells and medium. For these reasons, current research has focused on innovative continuous reactors such as the NASA microgravity bioreactor (7), hollow fiber reactors (8, 9), a single‐pass, plug flow reactor (10), a depth filter perfusion system (11), and spin‐filter bioreactors (12, 13). Despite continuing advances in cell culture apparatus technology, roller bottles are still prevalent throughout the industry because of their simplicity and relative effectiveness.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Mixing and Mass Transfer in a Recirculation Loop Results in High Cell Densities in a Roller Bottle Reactor

Berson¹,

Pieczynski²,

Svihla³

et al. 2002

Biotechnology Progress

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A recirculation loop added to a large-scale roller bottle reactor resulted in high cell densities as compared to standard roller bottles. Four different mammalian cell lines reached an average maximum density equal to 5.4 x 10(6) cells /mL (sigma = 0.263), which was between 2.13 and 2.95 times greater than the densities in roller bottles without recirculation using the same cell lines. The high densities were maintained over long durations (>25 days) while the reactor operated with continuous perfusion. The increased densities are attributed to enhanced liquid mixing and oxygen transfer that occur as a result of the recirculation loop. Models were developed that describe axial liquid flow and oxygen transfer in both the sample loop and the reactor growth chamber. Axial dispersion and oxygen transfer coefficients are presented for a variety of operating conditions. The increased oxygen transfer characteristics of the reactor allow for easy scale-up of roller bottle cultures by operating at larger volumes with greater liquid depths than conventional roller bottles permit. The surface-area-to-volume ratio in the tests performed was 0.206 versus 1.16 cm(-1) in a standard roller bottle.

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Development of a single‐pass ceramic matrix bioreactor for large‐scale mammalian cell culture

Cited by 10 publications

References 15 publications

Cyclic Operation of Ceramic‐Matrix Animal Cell Bioreactors for Controlled Secretion of an Endocrine Hormone. A Comparison of Single‐Pass and Recycle Modes of Operation

Cyclic Operation of Ceramic‐Matrix Animal Cell Bioreactors for Controlled Secretion of an Endocrine Hormone. A Comparison of Single‐Pass and Recycle Modes of Operation

A Self-Feeding Roller Bottle for Continuous Cell Culture

Enhanced Mixing and Mass Transfer in a Recirculation Loop Results in High Cell Densities in a Roller Bottle Reactor

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