A Controlled Environment System for Study of Mammalian Cell Physiology*

McLimans, William F.; Mount, David T.; Bogitch, Seymour; Crouse, Elizabeth J.; Harris, George F.; Moore, George E.

doi:10.1111/j.1749-6632.1966.tb41195.x

Annals of the New York Academy of Sciences

1966

DOI: 10.1111/j.1749-6632.1966.tb41195.x

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A Controlled Environment System for Study of Mammalian Cell Physiology*

William F. McLimans

David T. Mount

Seymour Bogitch

et al.

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1968

1973

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Cited by 25 publications

(2 citation statements)

References 47 publications

(10 reference statements)

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“…Prior 'reports have stressed the role played by cell dissociation ( 1 1 ), culture surface (12), kinetics of gas diffusion (13,14), and physiologic steady-state in the successful culture of differentiated liver cells (11,15,16). This preliminary report demonstrates the levels of hormone elaboration in thin-film culture systems of cells of endocrine origin under conditions similar to those employed in the cultures of liver cells.…”

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confidence: 77%

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Studies on Hormone Production by Human Fetal Pituitary Cell Cultures

Nussbaum

et al. 1970

Experimental Biology and Medicine

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confidence: 77%

“…The technique of cell cultures employed was described in detail previously (11). The pH control was achiteved by continuous gassing with 2 o'r 5% C02 in air with the appropriate bicarbonate level incorporated in the medium (12). The cell inoculum achieved was of heterogeneous composition, i.e., discrete cells and cell clumps.…”

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confidence: 99%

Studies on Hormone Production by Human Fetal Pituitary Cell Cultures

Nussbaum

et al. 1970

Experimental Biology and Medicine

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A pilot plant for mammalian cell culture

Moore

Hasenpusch

Gerner

et al. 1968

Biotech & Bioengineering

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SummaryStudies of the possible viral etiology of human leukemia have required large quantities of cultured cells derived from human hematopoietic tissues. Since cultures sufficiently large and free from contamination could not readily be produced according to existing methods, a pilot cell culture plant has been constructed for the production of mammalian cells in mass quantity. 500-ml to 20-liter trophocell units have already proved to be scientifically and economically practical, as they provide good reliability, excellent growth rates, and sustained yield cf human cells. 200-liter stainless steel culture units have now been added to the trophocell system. Five complete 200 liter units are now in operation. The design of the original stainless steel unit was based on that of a stainless steel, jacketed soup kettle. There are no openings in the vessel other than those in the lid, which provide convenient access points for sampling, sensor probes, etc. Environmental parameters, e.g., liquid level, temperature, and pH, are monitored and controlled with commercially available apparatus. Many initial problems connected with the new 200 liter units have been resolved, but operational and design problems remain in the areas of stable instrumentation, cell harvesting, salvaging and reuse of unspent media components, establishment of physiologic steady state, recovery of virus-containing cells with reculture of the remaining unaffected cells, and the recovery and separation of cell components and special products such as immunoglobulins, interferons, and hormones. A definitive cell plant with culture units of 20,50,250, and 1250 liters is now being constructed.We are impressed by the accumulating evidence of the divergent variations and alterations of cells in culture-morphologic as well as functional, by the extreme selective stresses imposed during the isolation and continuous culture of the mammalian cell in as well as by the ever-changing transient physiologic states encountered in the "batch type" or conventional cell culture ~y s t e m s .~ Of further importance is the increasing evidence that it may be feasible 625

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Rocker cell culture incubator

Tunnah

McLimans

Moore

1968

Biotech & Bioengineering

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SummaryA rocker mammalian cell culture system is described. The basic mechanical component is a special incubator with contained rocker culture trays. The system provides for precise environmental temperature regulation with intermittent rocking of contained cultures in such a manner as to permit thin fluid overlay of cells with maximum accessibility to oxygen while circumventing the problems of evaporation, substrate depletion, etc. IntroductionSeveral reports have delineated the critical role of gas diffusion in cell cultureOn the basis of experimental* considerations it has been demonstrated that good growth of human embryonic cells can be achieved if the culture fluid overlay is restricted to 0.5 mm or less. The fluid overlay depth of 1.5-5.0 mm utilized in mmt conventional cell culture systems severely limits access of oxygen to primary cells and in time creates an anoxic environment.3 Via the described rocker system, thin-layer culture waa achieved and demonstrated to be an important parameter in culture of cells representative of the tissue of origin.3The Rocker Culture SystemThe rocker culture system design is such as to provide a very thin layer of culture fluid over the mammalian cells. This establishes optimum conditions for rapid diffusion of gsses and thus ready access of oxygen to the cells. Intermittently, the thin fluid layer is replaced by a gentle rocking motion that permits reservoir fluid to bathe the cells. This effectively circumvents the problems of evaporation, exhaustion of substrate, and loss of pH control as anticipated in thin-layer culture systems incubated in the horizontal position. The Rocker IncubatorThe rocker incubator was designed on the basis of the following criteria: to provide a maximum number of cultures per unit space with minimum contact of the culture flask surface with the holding device; to provide ready access t o an individual culture flask ; to provide temperature-control circuits compatible with rocker incubator box providing a capability of temperature adjustment from ambient to 45°C and to control at the selected value within fO.1"C; to provide for rapid temperature recovery following biologic manipulative procedures ; to provide maximum reliability over long periods of time with minimum adjustment and maintenance ; to provide an electronically controlled drive mechanism BIOTECHNOLOGY AND BIOENGINEERING, VOL. X, ISSUE 5 X (1968) capable of rocking contained culture trays ~-2 5 ' off the level position with a smooth movement at a constant rate such as to prevent "sloshing" of culture fluid and, finally, to provide ready accesa for connections to auxiliary unita such as monitor flasks,' gas monitor,5 etc. COMMUNICATIONS TO THE EDITORSThree small incubators were constructed to allow concurrent investigations at different environmental temperatures as well as to decrease the temperature gradients usually encountered in large incubators.The incubators were designed to fit in a Par-Metal rack S o . PR-ii27-B vertically (Fig. 1). Plexiglas was chosen for favorable...

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A Controlled Environment System for Study of Mammalian Cell Physiology*

Cited by 25 publications

References 47 publications

Studies on Hormone Production by Human Fetal Pituitary Cell Cultures

Studies on Hormone Production by Human Fetal Pituitary Cell Cultures

A pilot plant for mammalian cell culture

Rocker cell culture incubator

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